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Sustainability 2010, 2(7), 2163-2175; doi:10.3390/su2072163
Article
Modeling of Metal Structure Corrosion Damage: A State of the Art Report
Department of Constructions and Mathematical Methods in Architecture, University of Naples Federico II, Via Forno Vecchio 36, 80134 Naples, Italy
* Author to whom correspondence should be addressed.
Received: 25 May 2010; in revised form: 18 June 2010 / Accepted: 5 July 2010 / Published: 15 July 2010
Download PDF Full-Text [370 KB, uploaded 15 July 2010 10:01 CEST]
Abstract: The durability of metal structures is strongly influenced by damage due to atmospheric corrosion, whose control is a key aspect for design and maintenance of both new constructions and historical buildings. Nevertheless, only general provisions are given in European codes to prevent the effects of corrosion during the lifetime of metal structures. In particular, design guidelines such as Eurocode 3 do not provide models for the evaluation of corrosion depth that are able to predict the rate of thickness loss as a function of different influencing parameters. In this paper, the modeling approaches of atmospheric corrosion damage of metal structures, which are available in both ISO standards and the literature, are presented. A comparison among selected degradation models is shown in order to evaluate the possibility of developing a general approach to the evaluation of thickness loss due to corrosion.
Keywords: atmospheric corrosion; metal and alloys; corrosion models
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Cite This Article
MDPI and ACS Style
Landolfo, R.; Cascini, L.; Portioli, F. Modeling of Metal Structure Corrosion Damage: A State of the Art Report. Sustainability 2010, 2, 2163-2175.
AMA Style
Landolfo R, Cascini L, Portioli F. Modeling of Metal Structure Corrosion Damage: A State of the Art Report. Sustainability. 2010; 2(7):2163-2175.
Chicago/Turabian Style
Landolfo, Raffaele; Cascini, Lucrezia; Portioli, Francesco. 2010. "Modeling of Metal Structure Corrosion Damage: A State of the Art Report." Sustainability 2, no. 7: 2163-2175.
Sustainability EISSN 2071-1050 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
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102 Bible Verses about Blood Of Jesus
1 John 1:7 ESV / 24 helpful votes
But if we walk in the light, as he is in the light, we have fellowship with one another, and the blood of Jesus his Son cleanses us from all sin.
Hebrews 9:22 ESV / 20 helpful votes
Indeed, under the law almost everything is purified with blood, and without the shedding of blood there is no forgiveness of sins.
Hebrews 9:14 ESV / 18 helpful votes
How much more will the blood of Christ, who through the eternal Spirit offered himself without blemish to God, purify our conscience from dead works to serve the living God.
Revelation 12:11 ESV / 12 helpful votes
And they have conquered him by the blood of the Lamb and by the word of their testimony, for they loved not their lives even unto death.
Acts 20:28 ESV / 12 helpful votes
Pay careful attention to yourselves and to all the flock, in which the Holy Spirit has made you overseers, to care for the church of God, which he obtained with his own blood.
Romans 5:9 ESV / 11 helpful votes
Since, therefore, we have now been justified by his blood, much more shall we be saved by him from the wrath of God.
1 John 1:7-9 ESV / 9 helpful votes
But if we walk in the light, as he is in the light, we have fellowship with one another, and the blood of Jesus his Son cleanses us from all sin. If we say we have no sin, we deceive ourselves, and the truth is not in us. If we confess our sins, he is faithful and just to forgive us our sins and to cleanse us from all unrighteousness.
Revelation 1:5 ESV / 8 helpful votes
And from Jesus Christ the faithful witness, the firstborn of the dead, and the ruler of kings on earth. To him who loves us and has freed us from our sins by his blood
Hebrews 13:20 ESV / 7 helpful votes
Now may the God of peace who brought again from the dead our Lord Jesus, the great shepherd of the sheep, by the blood of the eternal covenant,
Colossians 1:20 ESV / 7 helpful votes
And through him to reconcile to himself all things, whether on earth or in heaven, making peace by the blood of his cross.
Revelation 7:14 ESV / 6 helpful votes
I said to him, “Sir, you know.” And he said to me, “These are the ones coming out of the great tribulation. They have washed their robes and made them white in the blood of the Lamb.
Revelation 5:9 ESV / 6 helpful votes
And they sang a new song, saying, “Worthy are you to take the scroll and to open its seals, for you were slain, and by your blood you ransomed people for God from every tribe and language and people and nation,
Hebrews 13:12 ESV / 6 helpful votes
So Jesus also suffered outside the gate in order to sanctify the people through his own blood.
Hebrews 10:29 ESV / 6 helpful votes
How much worse punishment, do you think, will be deserved by the one who has spurned the Son of God, and has profaned the blood of the covenant by which he was sanctified, and has outraged the Spirit of grace?
Hebrews 10:19 ESV / 6 helpful votes
Therefore, brothers, since we have confidence to enter the holy places by the blood of Jesus,
1 Peter 1:18-19 ESV / 5 helpful votes
Knowing that you were ransomed from the futile ways inherited from your forefathers, not with perishable things such as silver or gold, but with the precious blood of Christ, like that of a lamb without blemish or spot.
Hebrews 10:6-9 ESV / 5 helpful votes
In burnt offerings and sin offerings you have taken no pleasure. Then I said, ‘Behold, I have come to do your will, O God, as it is written of me in the scroll of the book.’” When he said above, “You have neither desired nor taken pleasure in sacrifices and offerings and burnt offerings and sin offerings” (these are offered according to the law), then he added, “Behold, I have come to do your will.” He does away with the first in order to establish the second.
Romans 6:11 ESV / 5 helpful votes
So you also must consider yourselves dead to sin and alive to God in Christ Jesus.
John 6:67-71 ESV / 5 helpful votes
So Jesus said to the Twelve, “Do you want to go away as well?” Simon Peter answered him, “Lord, to whom shall we go? You have the words of eternal life, and we have believed, and have come to know, that you are the Holy One of God.” Jesus answered them, “Did I not choose you, the Twelve? And yet one of you is a devil.” He spoke of Judas the son of Simon Iscariot, for he, one of the Twelve, was going to betray him.
John 6:50-71 ESV / 5 helpful votes
This is the bread that comes down from heaven, so that one may eat of it and not die. I am the living bread that came down from heaven. If anyone eats of this bread, he will live forever. And the bread that I will give for the life of the world is my flesh.” The Jews then disputed among themselves, saying, “How can this man give us his flesh to eat?” So Jesus said to them, “Truly, truly, I say to you, unless you eat the flesh of the Son of Man and drink his blood, you have no life in you. Whoever feeds on my flesh and drinks my blood has eternal life, and I will raise him up on the last day. ...
Leviticus 17:11 ESV / 5 helpful votes
For the life of the flesh is in the blood, and I have given it for you on the altar to make atonement for your souls, for it is the blood that makes atonement by the life.
Genesis 9:4 ESV / 5 helpful votes
But you shall not eat flesh with its life, that is, its blood.
Hebrews 10:4 ESV / 4 helpful votes
For it is impossible for the blood of bulls and goats to take away sins.
2 Timothy 2:21-26 ESV / 4 helpful votes
Therefore, if anyone cleanses himself from what is dishonorable, he will be a vessel for honorable use, set apart as holy, useful to the master of the house, ready for every good work. So flee youthful passions and pursue righteousness, faith, love, and peace, along with those who call on the Lord from a pure heart. Have nothing to do with foolish, ignorant controversies; you know that they breed quarrels. And the Lord's servant must not be quarrelsome but kind to everyone, able to teach, patiently enduring evil, correcting his opponents with gentleness. God may perhaps grant them repentance leading to a knowledge of the truth, ...
Colossians 1:14 ESV / 4 helpful votes
In whom we have redemption, the forgiveness of sins.
Ephesians 6:10 ESV / 4 helpful votes
Finally, be strong in the Lord and in the strength of his might.
Ephesians 2:13 ESV / 4 helpful votes
But now in Christ Jesus you who once were far off have been brought near by the blood of Christ.
2 Corinthians 3:6 ESV / 4 helpful votes
Who has made us competent to be ministers of a new covenant, not of the letter but of the Spirit. For the letter kills, but the Spirit gives life.
1 Corinthians 10:15-17 ESV / 4 helpful votes
I speak as to sensible people; judge for yourselves what I say. The cup of blessing that we bless, is it not a participation in the blood of Christ? The bread that we break, is it not a participation in the body of Christ? Because there is one bread, we who are many are one body, for we all partake of the one bread.
Romans 10:17 ESV / 4 helpful votes
So faith comes from hearing, and hearing through the word of Christ.
Romans 8:1 ESV / 4 helpful votes
There is therefore now no condemnation for those who are in Christ Jesus.
Romans 3:25 ESV / 4 helpful votes
Whom God put forward as a propitiation by his blood, to be received by faith. This was to show God's righteousness, because in his divine forbearance he had passed over former sins.
Romans 2:4 ESV / 4 helpful votes
Or do you presume on the riches of his kindness and forbearance and patience, not knowing that God's kindness is meant to lead you to repentance?
Acts 4:12 ESV / 4 helpful votes
And there is salvation in no one else, for there is no other name under heaven given among men by which we must be saved.”
John 15:3 ESV / 4 helpful votes
Already you are clean because of the word that I have spoken to you.
Matthew 26:28 ESV / 4 helpful votes
For this is my blood of the covenant, which is poured out for many for the forgiveness of sins.
Matthew 9:6-8 ESV / 4 helpful votes
But that you may know that the Son of Man has authority on earth to forgive sins”—he then said to the paralytic—“Rise, pick up your bed and go home.” And he rose and went home. When the crowds saw it, they were afraid, and they glorified God, who had given such authority to men.
1 John 4:20-21 ESV / 3 helpful votes
If anyone says, “I love God,” and hates his brother, he is a liar; for he who does not love his brother whom he has seen cannot love God whom he has not seen. And this commandment we have from him: whoever loves God must also love his brother.
Ephesians 1:7 ESV / 3 helpful votes
In him we have redemption through his blood, the forgiveness of our trespasses, according to the riches of his grace,
1 Corinthians 6:19-20 ESV / 3 helpful votes
Or do you not know that your body is a temple of the Holy Spirit within you, whom you have from God? You are not your own, for you were bought with a price. So glorify God in your body.
Romans 3:23 ESV / 3 helpful votes
For all have sinned and fall short of the glory of God,
Acts 16:30-31 ESV / 3 helpful votes
Then he brought them out and said, “Sirs, what must I do to be saved?” And they said, “Believe in the Lord Jesus, and you will be saved, you and your household.”
Acts 11:14 ESV / 3 helpful votes
He will declare to you a message by which you will be saved, you and all your household.’
Acts 2:38 ESV / 3 helpful votes
And Peter said to them, “Repent and be baptized every one of you in the name of Jesus Christ for the forgiveness of your sins, and you will receive the gift of the Holy Spirit.
John 20:19-24 ESV / 3 helpful votes
On the evening of that day, the first day of the week, the doors being locked where the disciples were for fear of the Jews, Jesus came and stood among them and said to them, “Peace be with you.” When he had said this, he showed them his hands and his side. Then the disciples were glad when they saw the Lord. Jesus said to them again, “Peace be with you. As the Father has sent me, even so I am sending you.” And when he had said this, he breathed on them and said to them, “Receive the Holy Spirit. If you forgive the sins of any, they are forgiven them; if you withhold forgiveness from any, it is withheld.” ...
John 10:10 ESV / 3 helpful votes
The thief comes only to steal and kill and destroy. I came that they may have life and have it abundantly.
John 6:60-64 ESV / 3 helpful votes
When many of his disciples heard it, they said, “This is a hard saying; who can listen to it?” But Jesus, knowing in himself that his disciples were grumbling about this, said to them, “Do you take offense at this? Then what if you were to see the Son of Man ascending to where he was before? It is the Spirit who gives life; the flesh is no help at all. The words that I have spoken to you are spirit and life. But there are some of you who do not believe.” (For Jesus knew from the beginning who those were who did not believe, and who it was who would betray him.)
John 6:55-59 ESV / 3 helpful votes
For my flesh is true food, and my blood is true drink. Whoever feeds on my flesh and drinks my blood abides in me, and I in him. As the living Father sent me, and I live because of the Father, so whoever feeds on me, he also will live because of me. This is the bread that came down from heaven, not like the bread the fathers ate and died. Whoever feeds on this bread will live forever.” Jesus said these things in the synagogue, as he taught at Capernaum.
John 6:53 ESV / 3 helpful votes
So Jesus said to them, “Truly, truly, I say to you, unless you eat the flesh of the Son of Man and drink his blood, you have no life in you.
John 6:47-50 ESV / 3 helpful votes
Truly, truly, I say to you, whoever believes has eternal life. I am the bread of life. Your fathers ate the manna in the wilderness, and they died. This is the bread that comes down from heaven, so that one may eat of it and not die.
John 6:44 ESV / 3 helpful votes
No one can come to me unless the Father who sent me draws him. And I will raise him up on the last day.
John 1:12 ESV / 3 helpful votes
But to all who did receive him, who believed in his name, he gave the right to become children of God,
Luke 22:20 ESV / 3 helpful votes
And likewise the cup after they had eaten, saying, “This cup that is poured out for you is the new covenant in my blood.
Matthew 22:37-40 ESV / 3 helpful votes
And he said to him, “You shall love the Lord your God with all your heart and with all your soul and with all your mind. This is the great and first commandment. And a second is like it: You shall love your neighbor as yourself. On these two commandments depend all the Law and the Prophets.”
Isaiah 35:3 ESV / 3 helpful votes
Strengthen the weak hands, and make firm the feeble knees.
Isaiah 1:19-20 ESV / 3 helpful votes
If you are willing and obedient, you shall eat the good of the land; but if you refuse and rebel, you shall be eaten by the sword; for the mouth of the Lord has spoken.”
2 Chronicles 7:14 ESV / 3 helpful votes
If my people who are called by my name humble themselves, and pray and seek my face and turn from their wicked ways, then I will hear from heaven and will forgive their sin and heal their land.
1 Chronicles 28:9-10 ESV / 3 helpful votes
“And you, Solomon my son, know the God of your father and serve him with a whole heart and with a willing mind, for the Lord searches all hearts and understands every plan and thought. If you seek him, he will be found by you, but if you forsake him, he will cast you off forever. Be careful now, for the Lord has chosen you to build a house for the sanctuary; be strong and do it.”
1 Samuel 15:23 ESV / 3 helpful votes
For rebellion is as the sin of divination, and presumption is as iniquity and idolatry. Because you have rejected the word of the Lord, he has also rejected you from being king.”
Joshua 2:18 ESV / 3 helpful votes
Behold, when we come into the land, you shall tie this scarlet cord in the window through which you let us down, and you shall gather into your house your father and mother, your brothers, and all your father's household.
Deuteronomy 12:23 ESV / 3 helpful votes
Only be sure that you do not eat the blood, for the blood is the life, and you shall not eat the life with the flesh.
Exodus 24:8 ESV / 3 helpful votes
And Moses took the blood and threw it on the people and said, “Behold the blood of the covenant that the Lord has made with you in accordance with all these words.”
Exodus 12:13 ESV / 3 helpful votes
The blood shall be a sign for you, on the houses where you are. And when I see the blood, I will pass over you, and no plague will befall you to destroy you, when I strike the land of Egypt.
Genesis 9:6 ESV / 3 helpful votes
“Whoever sheds the blood of man, by man shall his blood be shed, for God made man in his own image.
Revelation 20:7-10 ESV / 2 helpful votes
And when the thousand years are ended, Satan will be released from his prison and will come out to deceive the nations that are at the four corners of the earth, Gog and Magog, to gather them for battle; their number is like the sand of the sea. And they marched up over the broad plain of the earth and surrounded the camp of the saints and the beloved city, but fire came down from heaven and consumed them, and the devil who had deceived them was thrown into the lake of fire and sulfur where the beast and the false prophet were, and they will be tormented day and night forever and ever.
Revelation 19:11-16 ESV / 2 helpful votes
Then I saw heaven opened, and behold, a white horse! The one sitting on it is called Faithful and True, and in righteousness he judges and makes war. His eyes are like a flame of fire, and on his head are many diadems, and he has a name written that no one knows but himself. He is clothed in a robe dipped in blood, and the name by which he is called is The Word of God. And the armies of heaven, arrayed in fine linen, white and pure, were following him on white horses. From his mouth comes a sharp sword with which to strike down the nations, and he will rule them with a rod of iron. He will tread the winepress of the fury of the wrath of God the Almighty. ...
1 John 4:14 ESV / 2 helpful votes
And we have seen and testify that the Father has sent his Son to be the Savior of the world.
1 John 2:2 ESV / 2 helpful votes
He is the propitiation for our sins, and not for ours only but also for the sins of the whole world.
1 John 2:1 ESV / 2 helpful votes
My little children, I am writing these things to you so that you may not sin. But if anyone does sin, we have an advocate with the Father, Jesus Christ the righteous.
1 Peter 2:9 ESV / 2 helpful votes
But you are a chosen race, a royal priesthood, a holy nation, a people for his own possession, that you may proclaim the excellencies of him who called you out of darkness into his marvelous light.
1 Peter 1:2 ESV / 2 helpful votes
According to the foreknowledge of God the Father, in the sanctification of the Spirit, for obedience to Jesus Christ and for sprinkling with his blood: May grace and peace be multiplied to you.
James 1:22 ESV / 2 helpful votes
But be doers of the word, and not hearers only, deceiving yourselves.
Hebrews 13:17 ESV / 2 helpful votes
Obey your leaders and submit to them, for they are keeping watch over your souls, as those who will have to give an account. Let them do this with joy and not with groaning, for that would be of no advantage to you.
Hebrews 10:19-20 ESV / 2 helpful votes
Therefore, brothers, since we have confidence to enter the holy places by the blood of Jesus, by the new and living way that he opened for us through the curtain, that is, through his flesh,
Hebrews 10:12 ESV / 2 helpful votes
But when Christ had offered for all time a single sacrifice for sins, he sat down at the right hand of God,
Hebrews 9:13-14 ESV / 2 helpful votes
For if the blood of goats and bulls, and the sprinkling of defiled persons with the ashes of a heifer, sanctify for the purification of the flesh, how much more will the blood of Christ, who through the eternal Spirit offered himself without blemish to God, purify our conscience from dead works to serve the living God.
Hebrews 1:3 ESV / 2 helpful votes
He is the radiance of the glory of God and the exact imprint of his nature, and he upholds the universe by the word of his power. After making purification for sins, he sat down at the right hand of the Majesty on high,
2 Timothy 2:15 ESV / 2 helpful votes
Do your best to present yourself to God as one approved, a worker who has no need to be ashamed, rightly handling the word of truth.
Colossians 2:1-23 ESV / 2 helpful votes
For I want you to know how great a struggle I have for you and for those at Laodicea and for all who have not seen me face to face, that their hearts may be encouraged, being knit together in love, to reach all the riches of full assurance of understanding and the knowledge of God's mystery, which is Christ, in whom are hidden all the treasures of wisdom and knowledge. I say this in order that no one may delude you with plausible arguments. For though I am absent in body, yet I am with you in spirit, rejoicing to see your good order and the firmness of your faith in Christ. ...
Ephesians 6:4 ESV / 2 helpful votes
Fathers, do not provoke your children to anger, but bring them up in the discipline and instruction of the Lord.
Galatians 5:13-14 ESV / 2 helpful votes
For you were called to freedom, brothers. Only do not use your freedom as an opportunity for the flesh, but through love serve one another. For the whole law is fulfilled in one word: “You shall love your neighbor as yourself.”
1 Corinthians 11:24-30 ESV / 2 helpful votes
And when he had given thanks, he broke it, and said, “This is my body which is for you. Do this in remembrance of me.” In the same way also he took the cup, after supper, saying, “This cup is the new covenant in my blood. Do this, as often as you drink it, in remembrance of me.” For as often as you eat this bread and drink the cup, you proclaim the Lord's death until he comes. Whoever, therefore, eats the bread or drinks the cup of the Lord in an unworthy manner will be guilty concerning the body and blood of the Lord. Let a person examine himself, then, and so eat of the bread and drink of the cup. ...
Romans 6:23 ESV / 2 helpful votes
For the wages of sin is death, but the free gift of God is eternal life in Christ Jesus our Lord.
Romans 3:24-25 ESV / 2 helpful votes
And are justified by his grace as a gift, through the redemption that is in Christ Jesus, whom God put forward as a propitiation by his blood, to be received by faith. This was to show God's righteousness, because in his divine forbearance he had passed over former sins.
Acts 15:17 ESV / 2 helpful votes
That the remnant of mankind may seek the Lord, and all the Gentiles who are called by my name, says the Lord, who makes these things
Acts 1:8 ESV / 2 helpful votes
But you will receive power when the Holy Spirit has come upon you, and you will be my witnesses in Jerusalem and in all Judea and Samaria, and to the end of the earth.”
John 17:21 ESV / 2 helpful votes
That they may all be one, just as you, Father, are in me, and I in you, that they also may be in us, so that the world may believe that you have sent me.
John 15:7 ESV / 2 helpful votes
If you abide in me, and my words abide in you, ask whatever you wish, and it will be done for you.
John 14:6 ESV / 2 helpful votes
Jesus said to him, “I am the way, and the truth, and the life. No one comes to the Father except through me.
John 8:24 ESV / 2 helpful votes
I told you that you would die in your sins, for unless you believe that I am he you will die in your sins.”
John 3:16 ESV / 2 helpful votes
“For God so loved the world, that he gave his only Son, that whoever believes in him should not perish but have eternal life.
John 1:29 ESV / 2 helpful votes
The next day he saw Jesus coming toward him, and said, “Behold, the Lamb of God, who takes away the sin of the world!
John 1:1 ESV / 2 helpful votes
In the beginning was the Word, and the Word was with God, and the Word was God.
Mark 1:15 ESV / 2 helpful votes
And saying, “The time is fulfilled, and the kingdom of God is at hand; repent and believe in the gospel.”
Zephaniah 2:3 ESV / 2 helpful votes
Seek the Lord, all you humble of the land, who do his just commands; seek righteousness; seek humility; perhaps you may be hidden on the day of the anger of the Lord.
Habakkuk 2:12 ESV / 2 helpful votes
“Woe to him who builds a town with blood and founds a city on iniquity!
Jeremiah 31:1-40 ESV / 2 helpful votes
“At that time, declares the Lord, I will be the God of all the clans of Israel, and they shall be my people.” Thus says the Lord: “The people who survived the sword found grace in the wilderness; when Israel sought for rest, the Lord appeared to him from far away. I have loved you with an everlasting love; therefore I have continued my faithfulness to you. Again I will build you, and you shall be built, O virgin Israel! Again you shall adorn yourself with tambourines and shall go forth in the dance of the merrymakers. Again you shall plant vineyards on the mountains of Samaria; the planters shall plant and shall enjoy the fruit. ...
Proverbs 4:23 ESV / 2 helpful votes
Keep your heart with all vigilance, for from it flow the springs of life.
1 Samuel 13:14 ESV / 2 helpful votes
But now your kingdom shall not continue. The Lord has sought out a man after his own heart, and the Lord has commanded him to be prince over his people, because you have not kept what the Lord commanded you.”
Deuteronomy 5:1-33 ESV / 2 helpful votes
And Moses summoned all Israel and said to them, “Hear, O Israel, the statutes and the rules that I speak in your hearing today, and you shall learn them and be careful to do them. The Lord our God made a covenant with us in Horeb. Not with our fathers did the Lord make this covenant, but with us, who are all of us here alive today. The Lord spoke with you face to face at the mountain, out of the midst of the fire, while I stood between the Lord and you at that time, to declare to you the word of the Lord. For you were afraid because of the fire, and you did not go up into the mountain. He said: ...
Hebrews 12:1-29 ESV / 1 helpful vote
Therefore, since we are surrounded by so great a cloud of witnesses, let us also lay aside every weight, and sin which clings so closely, and let us run with endurance the race that is set before us, looking to Jesus, the founder and perfecter of our faith, who for the joy that was set before him endured the cross, despising the shame, and is seated at the right hand of the throne of God. Consider him who endured from sinners such hostility against himself, so that you may not grow weary or fainthearted. In your struggle against sin you have not yet resisted to the point of shedding your blood. And have you forgotten the exhortation that addresses you as sons? “My son, do not regard lightly the discipline of the Lord, nor be weary when reproved by him. ...
Hebrews 10:1-39 ESV / 1 helpful vote
For since the law has but a shadow of the good things to come instead of the true form of these realities, it can never, by the same sacrifices that are continually offered every year, make perfect those who draw near. Otherwise, would they not have ceased to be offered, since the worshipers, having once been cleansed, would no longer have any consciousness of sins? But in these sacrifices there is a reminder of sins every year. For it is impossible for the blood of bulls and goats to take away sins. Consequently, when Christ came into the world, he said, “Sacrifices and offerings you have not desired, but a body have you prepared for me; ...
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19 Bible Verses about Wicca
Revelation 21:8 ESV / 6 helpful votes
But as for the cowardly, the faithless, the detestable, as for murderers, the sexually immoral, sorcerers, idolaters, and all liars, their portion will be in the lake that burns with fire and sulfur, which is the second death.”
Ephesians 5:11 ESV / 5 helpful votes
Take no part in the unfruitful works of darkness, but instead expose them.
Romans 1:25 ESV / 5 helpful votes
Because they exchanged the truth about God for a lie and worshiped and served the creature rather than the Creator, who is blessed forever! Amen.
Exodus 22:18 ESV / 5 helpful votes
“You shall not permit a sorceress to live.
Leviticus 19:31 ESV / 4 helpful votes
“Do not turn to mediums or necromancers; do not seek them out, and so make yourselves unclean by them: I am the Lord your God.
Leviticus 19:26 ESV / 4 helpful votes
“You shall not eat any flesh with the blood in it. You shall not interpret omens or tell fortunes.
John 14:6 ESV / 3 helpful votes
Jesus said to him, “I am the way, and the truth, and the life. No one comes to the Father except through me.
Deuteronomy 18:10-12 ESV / 3 helpful votes
There shall not be found among you anyone who burns his son or his daughter as an offering, anyone who practices divination or tells fortunes or interprets omens, or a sorcerer or a charmer or a medium or a necromancer or one who inquires of the dead, for whoever does these things is an abomination to the Lord. And because of these abominations the Lord your God is driving them out before you.
Exodus 20:1-26 ESV / 3 helpful votes
And God spoke all these words, saying, “I am the Lord your God, who brought you out of the land of Egypt, out of the house of slavery. “You shall have no other gods before me. “You shall not make for yourself a carved image, or any likeness of anything that is in heaven above, or that is in the earth beneath, or that is in the water under the earth. You shall not bow down to them or serve them, for I the Lord your God am a jealous God, visiting the iniquity of the fathers on the children to the third and the fourth generation of those who hate me, ...
Hebrews 9:27 ESV / 2 helpful votes
And just as it is appointed for man to die once, and after that comes judgment,
Ephesians 6:12 ESV / 2 helpful votes
For we do not wrestle against flesh and blood, but against the rulers, against the authorities, against the cosmic powers over this present darkness, against the spiritual forces of evil in the heavenly places.
Galatians 5:19-21 ESV / 2 helpful votes
Now the works of the flesh are evident: sexual immorality, impurity, sensuality, idolatry, sorcery, enmity, strife, jealousy, fits of anger, rivalries, dissensions, divisions, envy, drunkenness, orgies, and things like these. I warn you, as I warned you before, that those who do such things will not inherit the kingdom of God.
2 Corinthians 6:14 ESV / 1 helpful vote
Do not be unequally yoked with unbelievers. For what partnership has righteousness with lawlessness? Or what fellowship has light with darkness?
Romans 6:23 ESV / 1 helpful vote
For the wages of sin is death, but the free gift of God is eternal life in Christ Jesus our Lord.
John 8:44 ESV / 1 helpful vote
You are of your father the devil, and your will is to do your father's desires. He was a murderer from the beginning, and has nothing to do with the truth, because there is no truth in him. When he lies, he speaks out of his own character, for he is a liar and the father of lies.
John 3:36 ESV / 1 helpful vote
Whoever believes in the Son has eternal life; whoever does not obey the Son shall not see life, but the wrath of God remains on him.
John 3:16 ESV / 1 helpful vote
“For God so loved the world, that he gave his only Son, that whoever believes in him should not perish but have eternal life.
Luke 22:31 ESV / 1 helpful vote
“Simon, Simon, behold, Satan demanded to have you, that he might sift you like wheat,
Psalm 34:14 ESV / 1 helpful vote
Turn away from evil and do good; seek peace and pursue it.
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User:Susan Y Chen
From OpenWetWare
Jump to: navigation, search
I am a new member of OpenWetWare!
Contents
Contact Info
Susan Y Chen (an artistic interpretation)
I work in the Your Lab at XYZ University. I learned about OpenWetWare from from Bioe140L, a class at UC Berkeley, and I've joined because for a lab class.
Education
• Year, PhD, Institute
• Year, MS, Institute
• Year, BS, Institute
Research interests
1. Interest 1
2. Interest 2
3. Interest 3
Publications
1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Paper1]
2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Paper2]
leave a comment about a paper here
3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Book1]
All Medline abstracts: PubMed HubMed
Useful links
Personal tools
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[29arg] A fable of the Phrygian Aesop, which is well worth telling.
This work is licensed under a Creative Commons Attribution-ShareAlike 3.0 United States License.
An XML version of this text is available for download, with the additional restriction that you offer Perseus any modifications you make. Perseus provides credit for all accepted changes, storing new additions in a versioning system.
load focus Introduction (John C. Rolfe, 1927)
load focus Latin (John C. Rolfe, 1927)
hideData/Identifiers
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Document URN: urn:cts:latinLit:phi1254.phi001.perseus-eng1
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[25] In the same way also there are good works that are obvious, and those that are otherwise can't be hidden.
This work is licensed under a Creative Commons Attribution-ShareAlike 3.0 United States License.
An XML version of this text is available for download, with the additional restriction that you offer Perseus any modifications you make. Perseus provides credit for all accepted changes, storing new additions in a versioning system.
load focus Greek (Brooke Foss Westcott, Fenton John Anthony Hort, 1885)
load focus Latin (Saint Jerome, Bible Foundation and On-Line Book Initiative)
hide Places (automatically extracted)
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Do MSN Live.com Search Reinclusion Requests Work?
Mar 6, 2007 • 8:10 am | (3) by | Filed Under Bing Search
A WebmasterWorld thread asks if reinclusion requests at Microsoft's Live.com Search works.
Like with Google reinclusion requests, some go heard and some go unheard.
For example, one WebmasterWorld member has yet to hear back from Microsoft and his site is still not indexed. But another just had his site reindexed at Live.com after a reinclusion request was submitted 3 months ago.
As of today one of my sites, the one MSNDude said they ware realeasing from the ban has appeared in MSN. It's got about 20 pages indexed so I guess it is a start!
So how do you submit a reinclusion request at Live.com?
According to this help document, you can email webspam@microsoft.com.
How do you know if your site is not listed in Live.com?
If you suspect that your website was removed from the MSN Search index, you can use MSN Search to see whether a particular web page is still there. To see whether your web page is still in the index, type the url:keyword followed by the web addressof your Web page. For example, type url:www.msn.com/worldwide.aspx, and then click Search.
To see which web pages on your website are indexed, type the site:keyword followed by the web address for your website. For example, type site:www.msn.com, and then click Search. Because the web crawlersthat index websites manage millions of pages, it's normal for some pages to move in and out of the index.
To request reinclusion:
We may remove a website from the index if the website was reported as spam. If you suspect that your website was incorrectly identified as spam, please send an e-mail message to webspam@microsoft.com.
I also hear this online form make work as well.
Forum discussion at WebmasterWorld.
Previous story: Microsoft Refunds adCenter Advertisers Too Much
blog comments powered by Disqus
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Tabke Predicts Major Google Algorithm Update By Years End
Sep 8, 2011 • 8:45 am | (18) by | Filed Under Google PageRank & Algorithm Updates
A WebmasterWorld thread has a short but to the point prediction by Brett Tabke, the founder of WebmasterWorld. Brett feels that Google will be pushing out a major algorithm update, the size of Florida from 2003, by the end of the year.
Brett said:
Larry Page has been at the helm coming up on six months. According to various reports, LP has put alot of faith back in the algo nerds. I look for a Florida like update on the organic side before the end of the year.
Brett has been following Google updates since before Google was even doing real updates, so he knows his stuff.
But do you expect something as significant as Florida to hit by the end of the year, given we just had Panda in February?
Look at the Google update history outline:
• Fritz (Summer 2003)
• Florida (November 2003)
• Austin Update (Jan 2004)
• The Sandbox (April 2004)
• Bourbon (May 2005)
• Big Daddy (October 2005)
• Supplemental (Jan 2007)
• Paid Links (October 2007)
• Dewey Update (Mar 2008)
• Vince / Brand (March 2009)
• May Day (May 2010)
• Caffeine (June 2010)
• Scraper Filter (Jan. 2011)
• Panda (February 2011)
Major, major updates, rarely ever happen twice a year but I guess anything is possible.
We know Google is looking to improve their scraper detection but would that be the size of Florida? I doubt it.
Anyway, what do you think?
Forum discussion at WebmasterWorld.
Previous story: Google Says Thank You To Top Forum Posters By Holding A Summit
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Place:Carroll, Maryland, United States
Watchers
source: Getty Thesaurus of Geographic Names
source: Family History Library Catalog
the text in this section is copied from an article in Wikipedia
Carroll County is a county located in the U.S. state of Maryland. In 2010, its population was 167,134. Its county seat is Westminster.
Carroll County is a part of the Baltimore-Washington Metropolitan Area. While predominantly rural, the county has become increasingly suburban in recent years.
Contents
History
the text in this section is copied from an article in Wikipedia
Carroll County was created in 1837 from parts of Baltimore and Frederick Counties, see Hundred (division). It was named for Charles Carroll of Carrollton (1737–1832), signer of the American Declaration of Independence.
During the American Civil War, the population of Carroll County was sharply divided between supporters of the Union and the Confederacy. In 1863, there were significant troop movements through the county as part of the Gettysburg campaign. On June 29, 1863, the cavalry skirmish known as Corbit's Charge was fought in the streets of Westminster, when two companies of Delaware cavalry attacked a much larger Confederate force under General J.E.B. Stuart.
In 2013 the Carroll County Board of Commissioners voted to make English the official language of the county.
Timeline
Date Event Source
1837 County formed Source:Red Book: American State, County, and Town Sources
1837 Land records recorded Source:Red Book: American State, County, and Town Sources
1840 First census Source:Population of States and Counties of the United States: 1790-1990
1840 No significant boundary changes after this year Source:Population of States and Counties of the United States: 1790-1990
Population History
source: Source:Population of States and Counties of the United States: 1790-1990
Census Year Population
1840 17,241
1850 20,616
1860 24,533
1870 28,619
1880 30,992
1890 32,376
1900 33,860
1910 33,934
1920 34,245
1930 35,978
1940 39,054
1950 44,907
1960 52,785
1970 69,006
1980 96,356
1990 123,372
Research Tips
External links
This page uses content from the English Wikipedia. The original content was at Carroll County, Maryland. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Topic > Crime and Justice
Statistics by Topic
Violence
A Directory of Education and Training Statistics, 2009 (cat no. 1136.0)
Australia at a Glance, 2008 (cat no. 1309.0)
Australian and New Zealand Standard Offence Classification (ANZSOC), 2011 (cat no. 1234.0)
Australian Women's Year Book, 1997 (cat no. 4124.0)
Baby Boomers in Queensland: A Profile of Persons Born 1946-1965, 2005 (cat no. 4149.3)
Community Safety, Tasmania, Oct 1998 (cat no. 4515.6)
Crime and Justice News, July 2011 to June 2012 (cat no. 4500.0)
Crime and Safety, Australia, Apr 2005 (cat no. 4509.0)
Crime and Safety, Australia: Supplementary National and Standard Tables, 1998 (cat no. 4509.0.40.001)
Crime and Safety, New South Wales, Apr 2008 (cat no. 4509.1)
Crime and Safety, Queensland, April 1995 (cat no. 4509.3)
Crime and Safety, South Australia, Oct 2000 (cat no. 4509.4)
Crime and Safety, Victoria, April 1995 (cat no. 4509.2)
Crime Victimisation, Australia, 2011-12 (cat no. 4530.0)
Defining the Data Challenge for Family, Domestic and Sexual Violence, 2013 (cat no. 4529.0)
Defining the data challenge for family, domestic and sexual violence: Summary, 2013 (cat no. 4529.0.00.001)
Directory of Family and Domestic Violence Statistics, 2011 (cat no. 4533.0)
General Social Survey, Australian Capital Territory, 2006 (cat no. 4159.8.55.001)
General Social Survey, New South Wales, 2006 (cat no. 4159.1.55.001)
General Social Survey, Northern Territory, 2006 (cat no. 4159.7.55.001)
General Social Survey, Queensland, 2006 (cat no. 4159.3.55.001)
General Social Survey, South Australia, 2006 (cat no. 4159.4.55.001)
General Social Survey, Tasmania, 2006 (cat no. 4159.6.55.001)
General Social Survey, Victoria, 2006 (cat no. 4159.2.55.001)
General Social Survey, Western Australia, 2006 (cat no. 4159.5.55.001)
General Social Survey: Data Reference Package, 2002 (cat no. 4159.0.55.001)
General Social Survey: States and Territories, 2010 (cat no. 4159.0.55.003)
General Social Survey: Summary Results, Australia, 2002 (cat no. 4159.0.55.006)
General Social Survey: Summary Results, Australia, 2010 (cat no. 4159.0)
General Social Survey: User Guide, Australia, 2010 (cat no. 4159.0.55.002)
In Focus: Crime and Justice Statistics, July 2012 (cat no. 4524.0)
Information Paper: Crime and Safety, Expanded CURF, Australia, 2002 (cat no. 4509.0.55.001)
Information Paper: National Crime Statistics, 1993 (cat no. 4511.0)
Information Paper: National Criminal Justice Statistical Framework, 2007 (cat no. 4525.0)
Information Paper: National Information Development Plan for Crime and Justice, 2005 (cat no. 4520.0)
Information Paper: Sexual Assault Information Development Framework, 2003 (cat no. 4518.0)
Measures of Australia's Progress, 2010 (cat no. 1370.0)
Measures of Australia's Progress: At A Glance, 2008 (cat no. 1383.0.55.002)
Measures of Australia's Progress: Summary Indicators, 2009 (cat no. 1383.0.55.001)
Microdata: Crime and Safety, Expanded CURF, Australia, 2005 (cat no. 4509.0.55.002)
Microdata: General Social Survey, Basic CURF, Australia, 2006 (cat no. 4159.0.30.001)
Microdata: General Social Survey, CURF, Australia, 2010 (cat no. 4159.0.30.003)
Microdata: General Social Survey, Expanded CURF, Australia, 2006 (cat no. 4159.0.30.002)
Microdata: National Aboriginal and Torres Strait Islander Social Survey, Expanded CURF, 2008 (cat no. 4720.0.55.001)
Microdata: Personal Safety Survey, Expanded CURF, Australia, 2005 (cat no. 4906.0.55.001)
Microdata: Women's Safety Survey, Basic CURF, Australia, 1996 (cat no. 4128.0.15.001)
National Aboriginal and Torres Strait Islander Social Survey, 2008 (cat no. 4714.0)
National Aboriginal and Torres Strait Islander Social Survey, Australia, 2002 (cat no. 4714.0.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Australian Capital Territory, 2002 (cat no. 4714.8.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Australia: Summary Booklet, 2008 (cat no. 4714.0.55.003)
National Aboriginal and Torres Strait Islander Social Survey, New South Wales, 2002 (cat no. 4714.1.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Northern Territory, 2002 (cat no. 4714.7.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Queensland, 2002 (cat no. 4714.3.55.001)
National Aboriginal and Torres Strait Islander Social Survey, South Australia, 2002 (cat no. 4714.4.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Tasmania, 2002 (cat no. 4714.6.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Victoria, 2002 (cat no. 4714.2.55.001)
National Aboriginal and Torres Strait Islander Social Survey, Western Australia, 2002 (cat no. 4714.5.55.001)
National Aboriginal and Torres Strait Islander Social Survey: Data Reference Package, 2002 (cat no. 4714.0.55.002)
National Aboriginal and Torres Strait Islander Social Survey: Users' Guide, 2008 (cat no. 4720.0)
National Regional Profile, 2006 to 2010 (cat no. 1379.0.55.001)
NSW State and Regional Indicators, Dec 2010 (cat no. 1338.1)
Personal Safety Survey, Australia: State Tables, 2005 (cat no. 4906.0.55.004)
Personal Safety Survey, Australia: User Guide, 2005 (cat no. 4906.0.55.003)
Personal Safety, Australia, 2005 (Reissue) (cat no. 4906.0)
Pocket Year Book, Australia, 2000 (cat no. 1302.0)
Population Survey Monitor, Nov 2000 (cat no. 4103.0)
Publications and Products Released in January 2002, Jan 2002 (cat no. 1102.0)
Queensland Year Book (Hard cover), 2001 (cat no. 1301.3)
Recent Developments in the Collection of Aboriginal and Torres Strait Islander Health and Welfare Statistics, 2005 (cat no. 4704.0.55.001)
Regional Indicators, South Australia, 1998 (cat no. 1314.4)
Sexual Assault in Australia: A Statistical Overview, 2004 (cat no. 4523.0)
South Australia's Baby Boomers: A Profile, 2003 (cat no. 4149.4.55.001)
South Australian Year Book, 1999 (cat no. 1301.4)
Statistical Trends, NSW, 2007 (cat no. 1338.1.55.001)
Statistics Victoria, Jun 2011 (cat no. 1100.2)
Technical Manual: Crime and Safety, Expanded CURF, Australia, 2005 (cat no. 4509.0.55.003)
Technical Manual: Personal Safety Survey, Expanded CURF, Australia, 2005 (cat no. 4906.0.55.002)
The Health and Welfare of Australia's Aboriginal and Torres Strait Islander Peoples, Oct 2010 (cat no. 4704.0)
The Health and Welfare of Australia's Aboriginal and Torres Strait Islander Peoples: Summary Booklet, 2005 (cat no. 4704.0.55.002)
User Manual: ABS Remote Access Data Laboratory (RADL), Mar 2006 (cat no. 1406.0.55.002)
User Manual: Responsible Use of ABS CURFs, Sep 2009 (cat no. 1406.0.55.003)
Women's Safety Australia, 1996 (cat no. 4128.0)
Women's Safety, Australia: User Guide, 1996 (cat no. 4129.0)
Year Book Australia on CD-ROM, 2006 (cat no. 1301.0.30.001)
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
4602.0 - Environmental Issues: People's Views and Practices, Mar 2000
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 28/11/2000
Page tools: Print Page Print All RSS Search this Product
MEDIA RELEASE
November 28, 2000
Embargoed: 11:30 AM (AEST)
160/2000
Media alert - ABS Environmental Report 4602.0 will be released tomorrow
The Australian Bureau of Statistics (ABS) will release Environmental Issues, People's Views and Practices (cat. no. 4602.0) tomorrow, Wednesday November 29.
This annual report contains information about household waste management - including recycling and disposal of hazardous waste - motor vehicle ownership and maintenance, and use of private and public transport.
Much of the data is comparable with previous publications going back to 1996 and state breakdowns are provided.
The publication will be boxed at the Federal Press Gallery. Other media wishing to receive a copy electronically please email Adam Leavesley: adam.leavesley@abs.gov.au.
A media release and a summary of the of the publication will be posted to the ABS website at 11.30am, www.abs.gov.au.
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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23 January 2013
Daily archive · 1 posts
Stories from 23 January 2013
Resources: Guide to Creating Facebook Pages with Impact
It's no small task for small organizations and activists to put together a strategic Facebook page, and navigating around Facebook's setup pages can be confusing. Originally published in Arabic, Social Media Exchange (SMEX) just released the English version of their booklet Creating Facebook Pages with Impact. Breaking down how to use Facebook pages for activists, it addresses both technical aspects and management strategies.
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Tell me more ×
Answers OnStartups is a question and answer site for entrepreneurs looking to start or run a new business. It's 100% free, no registration required.
I've been running a nice dating site for the last 4 months and it's growing quite well and hope to be in alexa top 100k by end of year. So far it's only me doing pretty much everything, but I want to take things to the next level, mostly I want to find a partner who would put in time in exchange for equity. My personal network is pretty small and it's hard to find somebody who was the drive and commitment that I would like.
Are there startup partner matchmaking sites that I possibly could utilize ?
share|improve this question
1 Answer
Attend startups events, conferences, and perhaps consider applying to a startup accelerator/incubator. There is a list of startup incubators here:
http://answers.onstartups.com/questions/13615/are-there-any-similar-firms-out-there-like-y-combinator
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Tell me more ×
Answers OnStartups is a question and answer site for entrepreneurs looking to start or run a new business. It's 100% free, no registration required.
A few years ago I had an idea for an online business, its going to operate as a freemium model and simply deal with information no tangible goods. since then I have been creating the site learning html, css, rails etc etc from knowing nothing (hence the few years bit).
My site still isn't completely ready but the free side is usable. I would like to deploy it soon and see if it can get some traction and implement the premium side a little later.
Ideally I would like to setup as a private limited, payments will eventually (hopefully) be taken through a payment gateway such as PayPal Website Payments Pro, Sage Pay or WorldPay Global Gateway linked to my business bank account.
Can anyone advise as to how to set up an online business in the UK as regards to the law, company registering, setting up a business bank account and any other bits which needs to be done?
Thanks very much for any help its much appreciated!
share|improve this question
2 Answers
up vote 2 down vote accepted
Off the top of my head legals and regulations wise:
• Your accountant can usually set up the Ltd company for you. (Or use a company formation agent if you prefer)
• VAT Registration if you expect turnover to exceed £70k (iirc) - easy to do, again your accountant will usually do this
• Bank account - most accountants seem to recommend Cater Allen (as they no doubt get a commission), good alternative is the co op.
• Insurance (Employer, employee and public liability insurance are legally required). You can get these packaged together, and doesn't cost a great deal.
None of which are actually needed to start trading online. I'd recommend just launching it, as a part time self-employed, taking payments personally and step up to a LTD co when you have proved it works. Apart from anything else you have a yearly overhead of roughly £2,000 a year to run a Ltd Co (including accountancy)
For your own sanity set up a separate account for business use - even if it's just a personal savings account with bank to take payments into and keep separate from personal things. You might prefer a "doing business as" account so you can use the trading name.
You will need to let the Revenue know (when you start taking income, or within 3 months) or they hit you with an auto £100 fine :
http://www.hmrc.gov.uk/selfemployed/register-selfemp.htm
For payments also take a look at http://www.nochex.com/ over Paypal - UK based, their terms are better, and they have a business option to entirely free you from the risk of card fraud. (They're quite selective who's eligible for that option though). Good to get started with until you outgrow them.
If you're in the UK, selling to the UK also be aware of the Advertising Standards Regs as they now apply to websites, Distance Selling regs and Ecommerce requirements which say what you must have on the site. See here:
http://www.bodlelaw.com/e-commerce/web-site-legal-healthcheck
share|improve this answer
If it's a serious business idea then it's always best to see an accountant concerning whether or not you actually incorporation or not. You have the option of trading as a sole trader, or a private limited company. The limited company has the advantage of separating the financial dealings of your company and yourself more clearly, but the disadvantage of additional administration.
We're UK based and the process for registering with Sage Pay is pretty horrific if you're small. The key issue here is what will be the average size of payment, that determines, to an extent, which payment provider(s) are best suited.
But when you say you're ready to deploy soon, is this business idea entirely untested? If so, I'd run some kind of test before worrying about the admin side. If it goes nowhere, you can save yourself a lot of time and money.
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3answers
121 views
Can I pay by company card for myself?
I myself run a limited company in the UK. The bank offers me a free Visa card only if I make any payments using the card. The problem is that sometimes I don't need to buy anything for the company at ...
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Connexions
Sections
You are here: Home » Content » To listen for specific information
About: To listen for specific information
Module by: Siyavula Uploaders. E-mail the author
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Name: To listen for specific information
ID: m22109
Language: English (en)
Subject: Arts
License: Creative Commons Attribution License CC-BY 3.0
Authors: Siyavula Uploaders (support@siyavula.org.za)
Copyright Holders: Siyavula Uploaders (support@siyavula.org.za)
Maintainers: Siyavula Uploaders (support@siyavula.org.za)
Latest version: 1.1 (history)
First publication date: Apr 18, 2009 7:20 am -0500
Last revision to module: Apr 18, 2009 7:22 am -0500
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Version: 1.1 Apr 18, 2009 7:22 am -0500 by Siyavula Uploaders
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Error!
Success!
Extension Methods, a Simple HowTo
0
kicks
Extension Methods, a Simple HowTo (Unpublished)
o, what exactly is an Extension Method? From the C# 3.0 Specs (found here) they are 'static methods that can be invoked using instance method syntax. In effect, extension methods make it possible to extend existing types and constructed types with additional methods.'.
Kicked By:
Drop Kicked By:
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Wikia
SRD:Extend Spell
Talk0
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this wiki
Revision as of 23:12, August 11, 2009 by Surgo (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
This material is published under the OGL
Extend Spell [Metamagic]Edit
BenefitEdit
An extended spell lasts twice as long as normal. A spell with a duration of concentration, instantaneous, or permanent is not affected by this feat. An extended spell uses up a spell slot one level higher than the spell’s actual level.
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Skip to main content Help Control Panel
Lost? Search this Naples Florida website...|Add our search|Login A+ A- 54.234.42.16
Business Directory « Collier County Business Directory «
VALET OIL + LUBE
Register with us in one easy step!
Add your Press Release or Company Profile
Collier County Florida Company Profile.
Give your opinion about this listing below:
912 INDIAN RIVER ST IMMOKALEE FL 34142
2010-11-15 16:54:13
I am a page for VALET OIL + LUBE, which is a company located at: 912 INDIAN RIVER ST in the town of IMMOKALEE with the zip code of 34142
I was registered by CONTRERAS ADAM J who is at 912 INDIAN RIVER ST in IMMOKALEE FL, zipcode 34142
Phone:
Website:
Email:
. Map of VALET OIL + LUBE - 912 INDIAN RIVER ST, IMMOKALEE, FL 34142
Contreras Adam J will be the Editor of the 'valet-oil-+-lube' page.
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VALET OIL + LUBE
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912 INDIAN RIVER ST
IMMOKALEE FL 34142
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"url": "josm.openstreetmap.de/ticket/2996",
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Modify
Opened 4 years ago
Closed 4 years ago
#2996 closed defect (fixed)
error, but no Message, if a relation is containing itself
Reported by: tobhaeg@… Owned by: team
Priority: major Component: Core
Version: latest Keywords: relation error member contains itself
Cc:
Description
i hab the problem, that JOSM doesn't upload my data, after i've edited some relations. After a hard piece of work, i've found a relation, which was containing itself as a member (Members of rlelation 167346: A,B,167346,D, type:route,route:bus).
I had these Problems for at least two cases. after i've deleted the Relation from the member list, everything was OK.
The main Problem:
JOSM doesn't show any message of the problem. It's only standing at "Upload Data" for a endless time.
If these Error appears, it should be shown some message, why it isn't uploading the Data.
Attachments (0)
Change History (3)
comment:1 follow-up: Changed 4 years ago by Gubaer
comment:2 in reply to: ↑ 1 Changed 4 years ago by tobhaeg@…
comment:3 Changed 4 years ago by Gubaer
• Resolution set to fixed
• Status changed from new to closed
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"warc_url": "http://josm.openstreetmap.de/wiki/Plugins?action=diff&version=482"
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Changes between Version 481 and Version 482 of Plugins
Ignore:
Timestamp:
Mar 3, 2012 10:03:53 AM (15 months ago)
Author:
jttt
Comment:
Use direct address (without redirect) for todo plugin (because ant task check-plugins have problem with https -> http redirect)
Legend:
Unmodified
Added
Removed
Modified
• Plugins
v481 v482
99https://raw.github.com/Gubaer/josm-contourmerge-plugin/deploy/dist/contourmerge.jar
1010https://raw.github.com/Gubaer/josm-scripting-plugin/deploy/dist/scripting.jar
11 https://github.com/downloads/Gnonthgol/JOSM-todo/todo.jar
11http://cloud.github.com/downloads/Gnonthgol/JOSM-todo/todo.jar
1212
1313Unmaintained:
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[Date Prev][Date Next][Thread Prev][Thread Next][Date Index][Thread Index]
Re: [tdf-discuss] Re: LibreOffice for Academic Work -- College/University
Marc,
So far I've avoided commenting on your request for two main reasons: I
was obliged to retire last February so am no longer an "academic" as
such and also it was not clear what you were meaning.
Let me say that I never had problems with LibO (since the addition of
free-motion paths into Impress) for academic work. True, there are
differences in the way LibO and MS manage insertion of bibliography,
citations, references and so on but so what? You just get used to the
different way of working.
I had issues - probably my fault for not working it out - getting text
to flow round a graphic in any shape other than a rectangle. That can
be a real bother. I also don't like the way floating text boxes drift
around not just the page but the whole document when format changes
are made - or even worse when sections are moved!
I also never managed to put a working spreadsheet into a text
document, though it is something I use only rarely. Finally, the real
bugbear I have is that you can't "quick-roll" a database on the fly.
Either the database has to exist before you start (even if empty) or
you are tied into one highly restricted way of working.
I do hope this helps in some way.
On 16/05/2012 06:28, Marc Paré wrote:
> Hi Jonathan,
>
> Le 2012-05-16 01:02, Jonathan Aquilina a écrit :
>> Bibliography creation as well as source citations are very very
>> important.
>>
>> Regards
>> Jonathan Aquilina
>>
>> On May 15, 2012, at 9:32 AM, Marc Paré wrote:
>>
>
> Thanks, I am looking for people who are using specific extensions in
> their academic work. Maybe I should post this on the users' list as
> well. I was hoping that some people who work in academia could chime
> in and say what extensions they were using specifically for use in
> academia.
>
> Cheers,
>
> Marc
>
>
--
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abxm03q's bookmarks
"The way to get things done is not to mind who gets the credit for doing them."
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"Three things in human life are important. The first is to be kind. The second is to be kind. And the third is to be kind."
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Linda's quote collection
I'm female and made my book on 24th December 2007.
My book as a pdf
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Quotation added by staff
Why not add this quote to your bookmarks?
Love touched her heart, and lo! It beats high, and burns with such brave hearts. Crawshaw, Richard
This quote is about love · Search on Google Books to find all references and sources for this quotation.
A bit about Crawshaw, Richard ...
Richard Crawshaw, Baron Crawshaw of Aintree, known as Dick Crawshaw (September 25, 1917July 16, 1986) was elected as a British Labour Party Member of Parliament in 1964 but left, or "defected" as the media reported to join the SDP in 1981. Crawshaw was a strong supporter of the Territorial Army and on one occasion voted against the annual Defence Estimates because they failed to make a large enough provision for it.
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It's easy! Just pick the product you like and click-through to buy it from trusted partners of Quotations Book. We hope you like these personalized gifts as much as we do.
Make and then buy your OWN fantastic personalized gift from this quote
A boy doesn't have to go to war to be a hero; he can say he doesn't like pie when he sees there isn't enough to go around. Howe, Edgar Watson
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212 - The Extra Degree
The one extra degree makes the difference. This simple analogy reflects the ultimate definition of excellence. Because it's the one extra degree of effort, in business and life, that can separate the good from the great. This powerful book by S.L. Parker and Mac Anderson gives great examples, great quotes and great stories to illustrate the 212° concept. A warning - once you read it, it will be hard to forget. Your company will have a target for everything you do ... 212°
Click here to buy this »
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It's easy! Just pick the product you like and click-through to buy it from trusted partners of Quotations Book. We hope you like these personalized gifts as much as we do.
Make and then buy your OWN fantastic personalized gift from this quote
Those who hope for no other life are dead even for this. Goethe, Johann Wolfgang Von
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A selection of more great products and gifts!
212 - The Extra Degree
The one extra degree makes the difference. This simple analogy reflects the ultimate definition of excellence. Because it's the one extra degree of effort, in business and life, that can separate the good from the great. This powerful book by S.L. Parker and Mac Anderson gives great examples, great quotes and great stories to illustrate the 212° concept. A warning - once you read it, it will be hard to forget. Your company will have a target for everything you do ... 212°
Click here to buy this »
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Friday, May 12, 2006
Iran Sought Advice in Pakistan on Attack
Kathy Gannon, Yahoo News:
Pakistan's former army chief says Iranian officials came to him for advice on heading off an attack on their nuclear facilities, and he in effect advised them to take a hostage — Israel. Retired Gen. Mirza Aslam Beg said he suggested their government "make it clear that if anything happens to Iran, if anyone attacks it — it doesn't matter who it is or how it is attacked — that Iran's answer will be to hit Israel; the only target will be Israel."
Since Beg spoke in an interview with The Associated Press, echoes of his thinking have been heard in Iran, though whether they result directly from his advice isn't known.
Mohammad Ebrahim Dehghani, an Iranian Revolutionary Guards commander, was quoted last week as saying that if "America does make any mischief, the first place we target will be Israel." The threat was disavowed the next day by Brig. Gen. Alireza Afshar, deputy to the chief of Iran's military staff, who said it was Dehghani's "personal view and has no validity as far as the Iranian military officials are concerned."
And on Tuesday, Israel's vice premier, Shimon Peres, warned that "Those who threaten to destroy are in danger of being destroyed."
In the AP interview that took place several weeks before these threats were exchanged, Beg said a delegation from the Iranian Embassy in Pakistan had come to his office in January, seeking advice as Western pressure mounted on Iran to abandon its nuclear effort. Beg said he offered lessons learned from his experience dealing with India's nuclear threat.
He said he told the Iranians, whom he did not identify, that Pakistan had suspected India of collaborating with Israel in planning an attack on its nuclear facilities. By then, Pakistan had the bomb too. But both countries had adopted a strategy of ambiguity, he said, and Pakistan sent an emissary to India to warn that no matter who attacked it, Pakistan would retaliate against India.
"We told India frankly that this is the threat we perceive and this is the action we are taking and the action we will take. It was a real deterrent," he recalled telling the Iranians.
He said he also advised them to "attempt to degrade the defense systems of Israel," harass it through the Hamas government of the Palestinian Authority and the Hezbollah movement in Lebanon, and put second-strike nuclear weapons on submarines.
Although analysts are divided on how soon Iran might have nuclear weapons, Beg said he is sure Iran has had enough time to develop them. But he insists the Pakistani government didn't help, even though he says former Prime Minister Benazir Bhutto once told him the Iranians offered more than $4 billion for the technology. READ MORE
Ephraim Asculai, a former senior official with the Israel Atomic Agency Commission, said he didn't think Beg's remarks reflected official Pakistani policy.
Asculai said he believed Iran learned more from Iraq than from Pakistan, recalling that as soon as the 1991 Gulf War broke out, Saddam Hussein fired missiles at Israel, even though it wasn't in the U.S.-led coalition fighting Iraq.
Beg became army chief of staff in 1988, a year after Pakistan confirmed CIA estimates that it had nuclear weapons capability. He served until 1991 and now runs his own think tank. He speaks freely and in detail about the nuclear issue, but many critical blank spots remain and the subject remains one of great sensitivity, clouded by revelations in 2004 that A.Q. Khan, who pioneered Pakistan's nuclear bomb, sold nuclear technology to Iran, Libya and North Korea.
The bigger picture has also changed radically. Pakistan is now a U.S. ally in the war on terrorism, and Asculai said "Pakistani government officials have often suggested that they would be willing to have ties with Israel under certain conditions."
In the AP interview, Beg detailed nearly 20 years of Iranian approaches to obtain conventional arms and then technology for nuclear weapons. He described an Iranian visit in 1990, when he was army chief of staff.
"They didn't want the technology. They asked: 'Can we have a bomb?' My answer was: By all means you can have it but you must make it yourself. Nobody gave it to us."
The United States imposed sanctions on Pakistan in 1990, suspecting it was developing a nuclear bomb. In 1998, confirmation came with Pakistan's first nuclear weapons tests.
Although Beg insisted his government never gave Iran nuclear weapons, Pakistan now acknowledges that Khan sold Iran centrifuges to produce weapons-grade uranium, though without his government's knowledge.
In a televised confession Khan insisted he acted without authorization in selling nuclear technology to Iran, Libya and North Korea, saying the proliferation took place between 1989 and 2000.
Khan has been pardoned by President Gen. Pervez Musharraf, and Pakistan has refused to hand him over to the United States or the U.N. nuclear watchdog agency for questioning.
According to Beg, Iran first sent emissaries to Pakistan in the latter years of its 1980-88 war with Iraq with a shopping list worth billions of dollars, mostly for spare parts for its air force. It offered in return to underwrite the development plan of Gen. Zia-ul Haq, then Pakistan's ruler.
"Gen. Zia did not agree," he said.
Much of what Beg says cannot be independently confirmed, and the U.N.'s International Atomic Energy Agency did not respond to repeated requests for comment on Beg's version of events.
Another angle on these early contacts comes from Tanvir Ahmed, Pakistan's ambassador to Iran from 1987-1989. He said he had a rare meeting with Iran's nuclear inner circle in January 1988.
"It was the only time I was allowed in the inner sanctum of the nuclear discussions. I was asked to a lunch. ... they wanted to know whether Pakistan would help them on the nuclear side. They never said they wanted nuclear weapons. They said they wanted to master the nuclear cycle," Ahmed recalled.
Ahmed said he told them it was unlikely, but promised to relay the request to Zia. He said Zia told him: "You gave them the right answer."
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Help Wikitravel grow by contributing to an article! Learn how.
San José (Costa Rica)
From Wikitravel
(Redirected from San Jose (Costa Rica))
Jump to: navigation, search
The center of San José
San José is the capital of Costa Rica.
[edit] Understand
San Jose, the capital of Costa Rica, is on a plateau in the Central Valley at 1200 m (3,700 ft) elevation. It is ringed by lush green mountains and valleys. The population of this city is probably half of the whole country. It contains the primary airport, the University of Costa Rica, the US' and other embassies and many museums, cultural venues, hotels, markets, etc. It is the hub of the country.
[edit] Climate
The weather in San Jose Costa Rica can be different at all times depending of the route of the winds and yearly seasons, some times in the later months of the year it is colder than the months at the middle. For many Costa Ricans the coolest temperatures in San Jose are equal to high (warm) temperatures for a Northern American Citizen or some one from a country with a snow season. When you view the size of Costa Rica, then you can understand why weather temperatures differ from one city to another across short distances, unless the Caribbean weather suffers a major event in the atmosphere. Because of its elevation San Jose is usually 70 to 80°F (21 to 27°C) though it can get chilly at night. The rainy season is from mid April through December.
[edit] Get in
[edit] By plane
The airport (SJO) is 17km or about 20 minutes by car from the center of San José.
There is a local bus stop outside the airport (on the other side of the multi-story car park which you see when you come out of arrivals). It costs less than 1 US$ and takes you right downtown. You can put your luggage in the storage area below on the bus. The cheapest option is taking the bus into downtown and get a taxi there for your final destination. When you need to go to the airport from downtown, you catch the Alajuela bus (takes you to the airport) at the Merced Park. It costs less than $1 and they leave every five minutes and it takes about 25 minutes to get to the airport. The airport bus stop is easy to see. The Best Airport Shuttle service company is Easy Ride Shuttle Service [1] as they offer Airport Shuttle Express from/to your Hotel to SJO Airport The taxis charge around 25 US Dollars to take you to the city, be sure to take one of the licensed reddish-orange taxis that say "Taxi Aeropuerto." There are many unlicensed taxi drivers who will charge you almost twice as much as Taxi Aeropuerto. The taxis gladly take Dollars, but the local bus only takes Colones and they would not be pleased to get a 10 000 Colones bill.
There is an ATM by the entrance to the departures that will give you both Colones and Dollars.
[edit] By bus
Buses from Mexico, El Salvador, Guatemala, Honduras, Nicaragua and Panama both, arrive to and leave from San José.
The Tica Bus [2] terminal is the most common choice for locals and foreigners alike when it comes to traveling around Central America and even Mexico. Please take note that it has recently been moved to the other end of town, near the Mercedes Tower. (Address: 200 meters north and 100 meters west of Torre Mercedes (Paseo Colón), in front of the Magisterio Nacional Mortuary)
King Quality [3] is a new choice available, their prices are considerably more expensive or cheaper than Tica Bus depending on the destination. There is also Transnica, note they don't have a website up, for information their phone number is (506)2223-4123.
Of course most local buses start or end here. There are several bus terminals in San José [4]. It is important to know which bus terminal serves your bus route. Bus stops are usually every few blocks in the city. Take always a taxi, when traveling with luggage.And it is highly likely to speak to you when you arrive.
[edit] By train
Trains have recently made a comeback in Costa Rica and, after being shut down for many years, several routes have been put back into service using second-hand equipment brought over from Spain and some very ancient wooden carraiges that look like they have been taken from a museum. Lines are mostly singe-track and level crossings have no lights or protection at all, which has led to several accidents. There's also no signalling. Overall it's an interesting experience if you have the time and it's the best way of getting to Heredia (a lot faster and more comfortable than the bus).
Timetables for all services are available at: http://www.trenurbano.co.cr/horario_tren_urbano.aspx
Heredia: on weekdays, trains run betwen San José and Heredia every half an hour in the mornings (6am-9am) and afternoons (3:30pm-8pm), leaving from Estación del Atlántico near the Parque Nacional. Some of these trains continue on to the UCR and U Latina in San Pedro. The 6pm departure from San José (returning at 7pm) is a big train, so you can almost always get a seat on this one.
Pavas, San Pedro and Curridabat: another line runs through the south of the city, stopping at Estación del Pacifico, Sábana and heading west into Pavas and eventually turning round in a fairly dangerous slum area in the middle of the hills. If you take it east, it stops across the road from Estación del Atlantico and then goes to the UCR, U Latina and Curridabat. Timetables are very limited, with just one train per hour early in the morning and in the evening on weekdays.
Belen: A new service to Belen (just south of the airport) started on 5th April, leaving from Estación del Pacifico. Services are approximately every half an hour between 6-8am and 4-8pm on weekdays only and take 35 minutes.
[edit] Get around
Taxis are generally cheap. All taxis should have a meter. The fare starts at 570 colones, and is 570 per kilometre. Conversion is about 510 colones per USD (fluctuates daily). A ride inside the city center will normally cost 580-2500 colones. Basically a couple dollars, which they will accept, will get you anywhere in the city. Be aware that it is close to useless to give a taxi driver an exact street address. You have to point out some well-known building, park or hotel close to where you are going. Often there are no street signs and addresses are difficult to find, so be sure you know where you are going or you could get lost very easily. If you are driving in Costa Rica (one may see vehicles from Mississippi, British Columbia, Panama, and other places) note that the traffic lights don't have the yellow border around them and can sometimes be difficult to see, the road network is well utilized by locals (to overcapacity) so don't expect to get anywhere fast, also motorcycles weave in and out of traffic. Keep in mind the pet peeve most tourists have with tico kindness: often times when a tico has no idea where a certain destination you may have had in mind is, he or she will simply direct you to a random location. Often times simply incomprehensible, these directions are a reflection on the cultural approach to kindness many Costa Ricans adopt. Service Car Rental [5] has good chart about driving distances in Costa Rica. Or use the online route planner from YourTravelmap.com [6].
Easy Ride Shuttle service offers a New Airport Express shuttle Services for only $20 (1-2 people) $10 each (3 or more passengers)shuttle runs 24 hours( from any hotel San Jose downtown to SJO airport and viceversa) CHEAPER THAN TAXI FARES.
• Costa Rica Executive Transportation Chauffeur Driven Luxury Transportation and Security Service. Providing Transportation to anywhere in Costa Rica, air charters, meet and greet services, airport transfers for all travelers to Costa Rica.
• TransportationinCostaRica.com offers different airport shuttle services from the San Jose Airport to all Costa Rica destinations: Toll free 1-888-324 -9326
The Bus system is reliable, comfortable, extensive, and very cheap. For instance it costs about $5 to travel from San Jose to the Nicoya Peninsula.
It is as well possible to get around by bicycle in San Jose. If you want to buy a bicycle you find stores in Calle 6 / Av. 5 (Coca Cola) or south of "Avenida Segunda" on the corner or Av.6 / Calle 4. In the south east corner of plaza Viquez you find a small bicycle store.
[edit][add listing] See
• Museo de Oro Precolombino (The Gold Museum), an underground museum below Plaza de la Cultura. Tel 2243-4202, museoro@racsa.co.cr, [7]. Tu-Su 10AM-4:30PM. Entrance fee $9 for foreigners (or $4.5 with a student ID card). The collection consists of 1600 pieces of Pre-Columbian gold work dating from 500 AD to 1500 AD. Information is given on the processing and making of the pieces as well as their social, cultural and religious meanings. At the same place you will also find The Numismatic Museum and The Temporary Exhibition Galleries. There is a nice museum shop and a tourist office at the entrance.
• Museo del Jade (The Jade Museum), Avenida 7-9, Calle 9-11 bis, Instituto Nacional de Seguros,. Tel 2287-6034. Mo-Fr 8:30AM-3:30PM, SA:9:00am to 1:00pm. Entrance: USD $9. The largest collection of precolumbian Jade in America. Furthermore they show how Jade was elaborated in the precolumbian aera. They offer a variety of other objects made of stones, bones, ceramics, shells,... Moreover you get an insight in the daily life of the people in the precolumbian era. If you only have time for visiting one museum and interested in precolumbian , this is the recommended choice, it is small, easy to get in, very representative of the many materials used and geographic zones habited before the European contact.
• Museo de los Niños (The children's museum), Antigua Penitenciaría (the old prison). Tel. 258-4929. M-F 9:30AM-3:30PM, Sa-Su 10AM-4PM. Entrance fee 600 Colones for adults and 300 for children. This is an edutainment museum, and it was designed for Costa Rica's children, all the exhibits are in Spanish only. Not recommended as a visit, except when the Auditorio Nacional is hosting a concert or art gallery.
• Museo Nacional, Calle 17 Avenida 2. Tel 257-1433. Tu-Su 8:30AM-4:30PM. Entrance fee 2000 Colones. The museum includes a large butterfly garden (With many morpho butterflies) and a collection of large stone spheres from the Diquis Valley near the Pacific Ocean, a permanent precolumbian exhibition, the barracks, the rooms of the army general and his family, and a couple of temporal exhibits at the time. The museum building is an old fort called Cuartel Bellavista, in this place the Army was symbolically abolished by then president Jose Figueres Ferrer on December 1st, 1948 after the last civil war and armed conflict in the country.
• Museo de Arte Costarricense, east end of Sabana Park. This used to be San Jose's main airport terminal back when La Sabana was the airport. Tel 222-7155. Tu-Sa 10:00-16:00, Su 10:00-14:00. Entrance fee $5 (students $3).
• Insect Museum at the Universidad de Costa Rica A very elegant collection of exotic bugs. Only a few dollars, but check the times when they are open.
• Museo de arte y diseño contemporáneo (MADC) Definitely the main institution in Costa Rica dedicated to the broadcasting of contemporary art. Centro Nacional de la Cultura, Antigua Fábrica Nacional de Licores. Avenida 3, calle 15/17. San José, Costa Rica. Tel: (506) 2257 7202 / 2257 9370 Fax: (506) 2257 8702. Info related to current exhibitions, schedules and admission fees can be found at their oficial website www.madc.ac.cr[8].
• Zoológico Simón Bolivar An almost hidden zoo in Barrio Amón, some of the most representative animals are available in this small zoo. There are many big cats, including a non native lion, the serpentarium is one of the most interesting spots, with colorful (and dangerous) snakes available.
[edit] Things to Do
The main downtown area is a bustling collection of well-laid out streets filled with bustling traffic and lined with eclectic, historic architecture. On the surface it is a gritty downtown area, but look inside and you'll find friendly people, quirky spots, and the historic side of San Jose that change your impression. A walking tour is the best way to see this area.
[edit] Tours
• Barrio Bird Walking Tours offers downtown San Jose tours in Spanish/English covering everything from local history, art, architecture, and the "local" side of San Jose life. Flexible scheduling and reasonable prices starting $15usd with some packages including yoga, massage, and/or lunch. [9].
• Costa Rica Golf Day Tours (Tours to Valle del Sol, Cariari and Marriott Los Suenos), (732) 917-0177, [10]. 7am-4pm. Valle del Sol:$110, Cariari $110, Marriott Los Suenos: $225. All prices include hotel pickup and green fees. edit
There are a lot of other tours and local events and doings in and from San José. Some include:
• 3 in One
• Butterfly farm
• Coffee farm
• Canopy tours
• Rafting* Rios Tropicales (Whitewater tours from San Jose and other areas of Costa Rica), [11]. edit
• Volcanoes
• Buses to volcano Poas leave from Parque La Merced at 08:30AM daily (2990 colones).
• Buses to volcano Irazú leave at 8AM daily on Av. 2 in front of the National Theater (50 km, 2 h, 4000 colones return) and return at 12:30PM from the National Park. Park entrance fee for foreigners only: $US10 / 5020 colones, a bit overpriced but the view is nice.
• La Paz Waterfall Gardens An hour away from San Jose you can find lovely trails through primary rainforest that take you past five beautiful waterfalls. The La Paz water fall gardens also offer a hummingbird gallery, serpentarium, frog exhibit and large butterfly observatory.
[edit] Street Life
• Every Thursday a dozens of young jugglers gather at the Parque Morazan and juggle together. Often with percussion music. It s a free event in public space. Normally you will be invited to play with them. Lovely experience if you like street art.
[edit] Yoga
• Yoga. Costa Rica is a top destination for yoga. In San Jose, check out Downtown Yoga near Parque Morazan, which offers accessibly priced yoga and hooping classes for all experience levels. [12]
[edit] Sports
• Valle del Sol, Santa Ana (In the Lindora area next to Forum Park.), (506) 2282-9222, [13]. edit
• TTCR Golf Tour, (San Jose), 866 448 3182, [14]. 7am-10am. Specialized golf transportation to/from San Jose area hotels and Marriott Los Suenos course. Reserve online for a $160 rate (2 person min.) including trasportation. edit
• Gym. Decent gym facilities can be found at the Spa Corobici (telephone: 231-5542) located behind the Hotel Corobici. The taxi ride from the airport is approximately $10 - $20 USD and entry into the gym is 5,300 CRC or $10 USD. The club has a good selection of free and machine weights as well as a cardio theater. The club also has an outdoor swimming pool for lap swimming, a jacuzzi tub, and a sauna.
[edit] Casinos
• Casinos Many hotels have gaming. The most famous for the single traveler is Casino del Rey.
[edit] Learn
Costa Rica in general, and San José in particular, is a great place to improve your Español. Many people can speak some English and there are many Spanish classes available, including at the Universidad de Costa Rica, as well as "immersion" classes in private homes.
• Costa Rican Language Academy, [15] 1 (866) 230 6361 (in the US) Tel: +(506) 2280-1685" The Costa Rican Language Academy (CRLA) is a private institution specializing in Spanish language instruction. CRLA is completely owned and staffed by Costa Ricans, so their immersion programs provide students with the opportunity to experience the language and culture of Costa Rica first-hand. They also provide additional Costa Rican cultural education, with on-site dance lessons, and Costa Rican cooking classes. They will make arrangements for pick-up, travel arrangements, and the very effective immersion-type language training takes place from Monday to Thursday, allowing the students to have a three-day weekend each week to explore the many interesting elements of Costa Rica. The facility is in a good neighborhood in San Jose, and there is a nice boutique hotel for guests and family visitors (the Hotel Boutique Jade) just around the corner.
• Maximo Nivel, [16], de la Farmacia la Bomba 75 m sur (San Pedro, San Jose), ☎ 2253-9220, [25]. 7am-8pm. Maximo Nivel offers small group, online and individual Spanish classes taught by certified Native Spanish instructors and tailored to fit your schedule and needs. Via our optional Tandem Conversation Program, our Spanish clients can join free conversation practice when we team them up with local residents who are studying in our intensive English programs. Also, we provide intensive TEFL/TESOL certification classes each month that provide you with the credentials necessary to get a job as an English teacher anywhere around the country or the globe.
• Comunicare Linguistic, Cultural & Community Service Center [17] is a non-profit association dedicated to teaching Spanish as a second language, to fostering a deeper understanding of Central America and to supporting community development and volunteer work. Offers Spanish immersion, homestays, service learning, Central American studies, educational and entertaining excursions. Phone (506)2281-0432 E-mail: info@comunicare-cr.com
• Babylon Idiomas [18] is located in San José and 3 more places in Costa Rica and offers a wide range of affordable and high quality Spanish courses with experienced native teachers. The schools are open all year-round and new courses for all levels start every Monday. They organize a lot of cultural and fun activities for free and the personal service is really good. Accommodation options such as family stay or shared flat are available at an additional cost.
• Intensa [19] With locations in San Jose, Alajuela and Escazu, offers conversational lessons as well as home stays.
• Coastal Spanish Institute (CSI): Spanish and Surf school in Tamarindo, Costa Rica. Homestay or residence available, along will many fun activities.
• Wayra Instituto De Español [20] Programs run from 1-4 weeks, there's an option to include a surf course, the school is in Tamarindo Beach.
• Natural Spanish [21] With ecological tours, cooking and dancing lessons included.
• Centro Panamericano De Idiomas [22] Locations in Heredia, Monteverde and Flamingo Beach. The Flamingo location offers a chance to obtain an Open Water Diver Certification
• ISLS, Institute for Spanish Language Studies, is a service for Spanish students wanting to make the most of their experience in Costa Rica. Not only do they have first class language institutions throughout the country they offer their travel services to students. Once you are in Costa Rica, ISLS offers many valuable services that will help make your visit an enjoyable one. Spanish language students receive help finding schools and access to many services while they are in school. Knowledge of the layout of the country and Capital city is important for travelers, Spanish language students, people thinking about retiring or investing, or anyone planning on spending an extended period of time in Costa Rica. ISLS offers a Student Guest House in San Jose, Costa Rica that is available to all of our students whether they are in a Spanish program or have already completed one. It is a great place to start out with when entering the country for the first time. It is also a great place to recoup, check some scores, clean your cloths and just relax. ISLS has put together a "Welcome to Costa Rica program" so that clients have the easiest possible transition into the Costa Rican culture. You will arrive into Costa Rica three days earlier than your planned start date for your trip. They provide airport pick-up, accommodations, valuable safety tips, Costa Rican transit information, a “home base” while you travel, and an opportunity to meet other travelers with similar destinations and interests. Feel free to visit their website: [23]
• Intercultura Language Center, Heredia and Sámara Beach, +506-2656-0127, [24]. M-F 7AM-7PM. Intercultura offers immersion Spanish programs with daily cultural activities such as Latin dance, cooking, films, yoga, and arts. Homestay or hotel stays are available, and volunteer programs are offered. Two beautiful campuses, one on the beach, one in the colonial city of Heredia, close to San José. Class size 1-6 students, All teachers have university degrees and teaching credentials. US university credit available. from $199 per week. edit
[edit][add listing] Buy
• Boutique Annemarie, located inside Hotel Don Carlos, is a nice souvenir shop. But don't buy your stamps here, they'll charge you an extra 40% for the "service".
• El Pueblo shopping mall has lots of small souvenir shops.
• Mora Books Is a used book store on the corner of First Avenue and Street 3 in down-town San Jose. They have a great number of guide books. They will buy, trade, or sell for cheaper than I´ve found in other countries.
• Mercado Artesania One of the best places for getting souvenirs and handcrafted products. This market is on 2nd Avenue by the National Museum. [25]
• The Costa Rica Cigar Store Hundreds of Hand made Costa Rica Cigars. Locations on Sabana Sur and Barrio Amon. [26]
• Feria Verde de Aranjuez organic famrers market every Saturday from 07 am - 1 pm in the Barrio Aranjuez [27]
The best coffees have deserved reputations for superb quality. Supermarkets/grocers and small coffee growers usually have better prices than shops that cater to tourists. Often packaged in 12 oz. sealed bags, you should only purchase roasted, whole beans rather than ground...for epicures, "strictly hard bean" (SHB)). They will keep flavor longer...until you can store them properly at home (Google for methods), and won't include sugar as often found in Costa Rican ground. Roasted coffee also prevents you from running afoul of agencies such as FDA/APHIS that requires special licensing for importing "green"/unroasted beans (may be considered plant material).
You will find a huge selection of Cuban Cigars at The Havana Humidor Room, 2 blocks North and 1 block West of the Holiday Inn.
Throughout the city, there is also a good deal of shops with wooden and ceramic souvenirs. The wooden pieces, such as masks, plaques, and other forms of wall art, are all beautifully hand carved as well as hand painted and the artisan usually signs their work with their name and where it was made on the backside. The ceramic pottery and dishware is done in this similar fashion and are available in a variety of designs and colors. These make interesting and personally unique gifts to bring home to family and friends for a reasonable price.
• Havana Humidor Room, Av. 9 y Calle 3.5 (2 blocks N. and 1 blk. W. of downtown Holiday Inn), 506 85282290, [28]. 11 to 7. Largest selection of genuine Cuban cigars in Central America edit
[edit][add listing] Eat
Mercado Central is a very old, interesting and bustling food market, which also contains a number of small restaurants and quick-serve counters for the locals. You will find fresh cooked fish and shellfish, corn based dishes, sopa de pescado (fish soup) and such exotics as "squid in his ink", ceviche (small bits of raw fish "cooked" in lime juice), helado de sorbetera (artesanal local cinnamon ice-cream) and more. Perhaps not for the faint of heart. And you could always just go for La Calle - Anything a street vendor is selling is probably good, for example the Mangos, street vendors often sell unripe mango strips with salt and lime, it's great.
• Bar Poas, Avenida 7, Calle 3 y 5 (Two doors down from the Pangea Hostel, across the street from St. Thomas Hotel in Barrio Amon), (506) 2-223-8677, [29]. 1200-0230. Photos of regular customers adorn the wall of this dimly lit but friendly bar and restaurant. A decent menu of Gringo and Tico food is available from noon to 0230. Try the award-winning Chili con Carne. There´s always an interesting group of characters there including proprietor Harry Hart who is always willing to dispense useful advice about San Jose and Costa Rica. edit
• Bagelmen's, (Escazú, San Pedro, Curridabat, Guachipelín, Belén, Heredia), [30]. Good prices, excellent quality fast and healthy food (bagels, cream cheeses, coffee, salads, desserts and more). Very popular among tourists and american expats. Free internet. edit
• Cafetería Parisien, (Gran Hotel, between Theatro Nacional and Plaza de Cultura). Not the most exciting food, but elegant settings. edit
• Ganesha, (Located in Hotel America, 50 meters south of Central park in Heredia), 8379-7951. This is an Indian, Mediterranean and Middle Eastern restaurant in Heredia, Costa Rica just about 15 minutes drive from the airport. Indian food lovers can get their garam masala fix here. Great Hummus, Baba Ganoush, Seekh Kebab and Chicken Tikka Masala. The lassis are great too edit
• Hotel Grano de Oro, Calle 30 Avenida 2/4, 255-3322. Beautiful but expensive restaurant. A breakfast menu costs around 2400 Colones or treat yourself to the delicious banana-macadamia nut pancakes (2000 Colones). The entrees are diverse and creative, do not hesitate to try the soup of the day, even if it sounds less than exciting. Duck is on the menu, and is some of the best to be had anywhere. Numerous seafood dishes are cooked to perfection, as is the beef and pork. edit
• Ristorante Tutti Li, Plaza Itskatzu, Escazú (Near Multiplaza, Courtyard, Holiday Inn, Residence Inn, Hotel Intercontinental), 2588-0425, [31]. One of the best Italian restaurants in town. Fresh pastas, traditional italian plates, brick oven pizza and exquisite wine selection, all with a modern flare. A must while in San José. Located in one of the most premium locations in the city. edit
• Hotel Vesuvio Restaurante, Barrio Otoya, 257-5411. Italian food, pastas 1900 Colones, chicken 2000-2500. edit
• Lukas, Centro Comercial El Pueblo. One of two other restaurants at the El Pueblo. edit
• Machu Picchu Restaurant, (Paseo Colón) 1st Ave (125 meters north from Kentucky FC restaurant), 506.222.3679. The best international Peruvian Food & Sea Food. Try the Causa Rellena, Cebiche, Lomo Saltado, Ají de Gallina and Peru's traditional and landmark drink: Pisco Sour. edit
• Pizzeria Il Pomodoro, Cerca de Parque Kennedy y Banco Nacional en San Pedro. One of the best known casual restaurants in Costa Rica, . Italian cuisine, very good pizza and pasta, good cheap wine, from second floor great view or the mountains to the west. edit
• Soda Esnider, (Walk a few blocks to the east from Plaza de la Cultura over the Central Avenue and get into Galerías Avenida Central. You will find several stores and if you go down the stairs you will come across Soda Esnider). You can have a wonderful “Casado” for lunch with a natural drink for as little as 1600 Colones ($3) edit
• Tin Jo, Paseo de los estudiantes. This is an incredible Asian restaurant featuring Japanese, Thai, Chinese and Indian food. Don't be scared by the looks on the outside, the restaurant is clean, beautifully decorated and offers the best service I experienced in Costa Rica. edit
• Vishnu. Vegetarian and Organic Restaurant. , With very reasonable prices and a varied, tasty menu they are easily the best choice for vegetarian food in SJ. There are three locations downtown San Jose: - 1 - South side boulevard, near the Banco popular. - 2 - Across Omni Center. - 3 - North side of the Iglesia Del Carmen. edit
• El Balcón del Marisco, appx 1 Km. East of EPA toward to hwy to Cartago. 10:00 a.m. - 11:00 p.m.. Great place for fish. Always busy. Service very good. No other place comparable. Clean and safe. ~$11.00. edit
[edit][add listing] Drink
• El Pueblo [32] Is a shopping center which becomes a clubbing district at night. There are several bars and nightclubs cluttered in the tiny little alleyways. Just mention El Pueblo to your taxi and he'll know where to go.
• San Pedro Is home to many bars and clubs, try to avoid the university of Costa Rica area at night as the many bars in the university district tend to attract brawls and even the occasional bullet shot. The other bars/clubs in San Pedro are free of trouble.
• Barrio La California Is the place to be if you're into the bohemian/rock crowd. Many bars have local bands playing every night. Great place to go bar hopping!
[edit] Gay and Lesbian nightlife
San Jose is a very tolerant city. Most bars are gay friendly. There is a small but vibrant gay life in San José from lesbian bars, to saunas and discos.
• La Avispa Is the oldest gay disco in Costa Rica. Big place with Latin music and dancing, pop. It has the most popular crowd and is recommended if you want to try the local flavor. Sunday's late afternoon and evening is the best time to go.
• Club OH Huge electronic music place with open bar on Fridays and Saturdays. It has a VIP area with better quality liquors and another DJ. Live Drag shows at midnight. Best day to go is Saturday.
• Bochinche Crowded Pop bar/disco, twink ambient. It is a very fashionable art deco style, colorful place. Diverse shows during the whole week, Fridays are open bar with carnival at midnight. Place is open until 5AM. Gets very crowded after 1AM. $3 each person.
[edit][add listing] Sleep
San Jose is full of hotels from one star to perhaps three. Closest to the airport are Hampton Inn, Doubletree and Marriott.
[edit] Budget
• Hostel Casa del Parque, On the northwest corner Parque Nacional (Avenida 3 and Calle 19) (Barrio La California), 506 2233 3437 (), [33]. checkin: 2pm; checkout: 11am. Dorms from $8. Comfortable beds with real mattresses in dorm style rooms of 13 beds. Five private rooms including one deluxe. Fully-quipped kitchen. There is also a common room equipped with cable TV, free internet, and coffee. Very central, tranquil location in an old but well-maintained house. Excellent hosts, knowledgeable and friendly. $8. (-84.069752,9.934538) edit
• Casa Ridgeway, C 15, Av 6/8, +506 2221-8288, [34]. A comfortable Quaker hotel with a shared kitchen and dining area available for use, a great place to meet like minded travelers, a simple breakfast is included. dorms, singles, doubles, triples available $10-$12 /person. edit
• Casa Yoses Hostel, Avenida 8, Calle 41, Los Yoses, 250 west from Spoon, +506 2234-5486 (), [35]. Casa Yoses is in a great central location surrounded by tons of restaurants, bars, and stores, just off the University of Costa Rica and the San Pedro Mall. edit
• Costa Rica Backpackers, Avenida 6, calle 21,23, +506 2221-6191 (), [36]. Great place to meet other travelers in town, 20 minutes walking from the center, affordable and comfortable hostel, facilities include an outdoor pool, kitchen, free internet and bar. Great place to party! Dorm : 12 USD. edit
• Galileo Hostel, Corner Calle 40, Avenida 2, 506.2248.2094 / (US)617.718.3531 (), [37]. Colorful hostel with a Caribbean vibe. Free internet and wifi, towels, common kitchen, in house bar, Wii, guitars, frequent theme nights, and hot showers. Close to many embassies and the largest public park in the city. New owners as of January 2009. Dorm $9-10 US, double room $22-30 US pp. edit
• Gaudys Backpackers Hostel, (Close to Parque La Sabana, 2km west of town), (), [38]. Cnr Ave 5/Calle 36-38, phone (506) 22480086, free breakfast, internet and WiFi, kitchen and laundry use, storerooms, dorms from USD 12. edit
• Hostel Bekuo, Avenida 8, Calle 40, Los Yoses, 325 West of Spoon Los Yoses, +506 2234-1091 (), [39]. Hostel Bekuo is one of the nicest and cleanest hostels in all of Costa Rica, and you can't ask for a better location while staying in San Jose. Includes free breakfast, pool table, internet, coffee & tea, and above all a great atmosphere and staff edit
• In & Basic Hostel Lounge, 300 m South and 75 m west of Spoon, Los Yoses. Fátima´s church North side, (Barrio Los Yoses, San Pedro), 506 2234 2998 (), [40]. Lounge Hostel with Private rooms (with/out baths, from 15 USD per person) and dorms (from 11 USD), Breakfest included. Free WiFi inside the rooms, pool table, air-hockey table, a garden with BBQ area, kitchen for free use. All new and modern. 2 blocks away from bars, supremarket and the San Pedro Mall. One hostel, One home, A different way of living in San Jose! edit
• JC & Friends Hostel, Cnr Ave 3/Calle 34, (506) 83748246. Phone , free breakfast, internet, kitchen use, dorms from USD 10. Near Tica Bus Terminal. edit
• Kabata Hostel, Avenida 9-11, Calle 7 #927, Downtown San Jose (Barrio Amón, 225 m North of kiosk park Morazán), +506 2283-2000, 2255-0355, 2255-3264 (), [41]. Kàbáta Hostel is centrally located and provides free breakfast, internet, and lots of helpful information. Dorm: $12, private: $32. edit
• Molino Rojo Hostel, Paseo Colón y Calle 32, diagonal a KFC, (506) 22216868 (), [42]. Brand new hostel in a safe and central area of San José with dorm beds from $10 and private rooms from $25 per night. Free breakfast, free use of the internet, communal kitchen, garden, lounge and bar. Big party the first Saturday of every month! edit
• Tranquilo Backpackers, Calle 7 esq. Avenida 9-11 (250 m North of park Morazán, in front of Kabata Hostel), (506) 2223-3189, 2222-2493 (), [43]. Clean and safe with a fun and lively atmosphere with guitars and movie everyday at 8PM. Free breakfast everyday, free coffee and tea all day, free internet/wifi, free storage, full kitchen, and a friendly knowledgeable staff. ATTENTION now closed Dorms from $10, singles from $17, doubles from $24. edit
• Hotel Danubio, Calle 18 Avenida 3 (75 m North of estacion Coca Cola), (506) 2221-9446. checkout: 1 PM. Clean and safe and extremely peaceful. You'll get a good nights sleep here without having to worry about all the hippies in CR, free internet/wifi, and a friendly knowledgeable staff. No Dorms, singles from $18, doubles from $25. edit
• '''Hostel Casa Colon''', Paseo Colon, C 24 North, in front of Torre Mercedes, right in the corner, +506 2256-0276 (), [44]. The number one and first Premium Hostel in Central America, at a Hostel prices, with high quality accomodations, Hostel located in San Jose Costa Rica in the best tourist-residential and quick access area in San Jose. Dorms, Private rooms,private and shared bathrooms, huge flat screen TVs, big lounge with wire/wireless high speed internet 24/7,video surveillance, International Restaurant and Cafe-Bar, airport shuttle service, free tourist info and travel tips, free maps, free parking, typical costarican breakfast included, plenty of services around the area, public and private transportation to anywhere in the country upon just a few steps, and much more! edit
• Castle Tam Hostel, 100 al oeste de la Escuela Roosevelt (san pedro san jose), 619 866 4553 (), [45]. checkin: 24 hours; checkout: 11:00am. $10 and $12 a night dormitory rooms. $20, $29 and $39 a day private rooms available. There are also monthly rates from $295, $370 and $495. Castle Tam is located in the safe San Pedro/Los Yoses area close to central San Jose and the University District of Universidad de Costa Rica. Its next to all the bus terminals, government agencies, cheap restaurants, museums and historical centers. Offers free WIFI, free city tours and pub crawls on student Calle de La Amargura From $10. (9.930908,-84.053763) edit
[edit] Mid-range
• Adventure Inn, Tel: +506-2239-2633, [46]. Only 8 minutes from the airport with free transportation. $77 single, $94 double, includes an all you can eat American breakfast, free airport transportation, free international phone calls, on-site sports bar / restaurant, pool and Jacuzzi, fully-equipped gym, hotel-wide wireless internet. Click here for a 10% discount off your room rates for cash or travelers cheques or mention WikiTravel at your time of booking.
• Airport Hotel, Tel: +506-24337333, [47]. Only 5 minutes from the international airport with free transportation 24 hours. Wikitravel corporate rate $70 single, $80 double, includes taxes & buffet breakfast, restaurant, pool and wireless internet. Mention WikiTravel when booking through our website.
• B&B Tambo Mundo, del Cristo de Sabanilla 400, tel. +506 2273 0265 / +506 8848 9707, juancarlos@tambomundocostarica.com, [48]. Tambo Mundo is a bed & breakfast with an amazing view of Braulio Carrillo National Park and Irazú Volcano in a big house with cozy common places. 5 rooms with plenty of hot water, delicious breakfast ( close to the University of Costa Rica and San Pedro. $30- $45 single occupation including taxes. A big Latin American literature, sustainable development and politics library, movies and documentaries are available. Fast Wireless Internet connection all around. Telephone and cable TV. Laundry service (free for long stays). Fluent English/Spanish. German and French.
• Costa Rica Guesthouse, tel. 506 2223 7034, [49]costaricaguesthouse@gmail.com. From $35 a night, located in downtown San Jose near the supreme court, housed in a beautifully restored 1904 building, this fine guesthouse features 23 decorated private rooms with king size beds, semi-orthopedic mattresses and free wifi access. Especially designed for couples, families and people looking for a little extra comfort, Costa Rica Guesthouse is the upscale hotel at budget price.
• Courtyard by Marriott San Jose, Autopista Prospero Fernandez, Calle Marginal N., Plaza Itskatzu San Jose, Tel:+ 506 2208 3000, [50] The Courtyard San Jose is located nearby San Jose's industrial parks, businesses, international companies and within minutes from shopping, nightlife plaza and the best restaurants in San Jose. Spacious rooms, free high-speed Internet, on-site restaurant, outdoor pool and health club.
• Hemingway Inn, Barrio Amon
Hemingway Inn, Avenida 9, Calle 9, Barrio Amon, (Behind I.N.S.), Tel +506 221-1804, reservations@hemingwayinn.com, [51]. Former mansion turned into a hotel. Friendly staff. Quiet neighborhood. Safe and secure. Decent price for the area: $35-45 for a double including traditional breakfast with Gallo Pinto and Tamales. Hotel has a small bar and hot tub on the premises. The hotel will organize Eco-tours for the rest of Costa Rica. Free Internet service and Wi-Fi.
• Hotel & Casino Europa, Central St. Between 3th & 5th Ave., (506) 2222-1222 (, fax: (506) 2221-3976), [52]. edit
• Hotel Del Rey, Tel: 1-866-765-8037, [53]. Infamous, overpriced, unsafe, and overrated. Sort of a decadent area (prostitution). You're better off hanging out in one of the many other, better priced places, no matter what your buddies have told you.
• Hotel La Rosa de America, Tel: +(506) 2433-2741, http://larosadeamerica.com/. Located 15 minutes west from the San Jose Airport, in the town of La Garita de Alajuela. This hotel offers sparking clean accommodation in twelve rooms and two family suites arranged in cabinas, located around a large beautifully maintained tropical garden and swimming pool. Breakfast at the restaurant[54] is included.
• '''Hotel Out of Bounds''' (Hotel Out of Bounds Escazu), Escazu, +506 2288 6762 (), [55]. 70$. edit
• Hotel Presidente [56]. Located in downtown San Jose, on the walking boulevard of Avenida Central and Calle 7 (seven street). Great location close to historic landmarks, museums and plazas. 4 star accommodations with rooms starting at $85 + tax per night. Free internet. Buffet breakfast included.
• Hotel San Gildar, next to the Costa Rica Country Club, [57]. A beautiful private hotel in the higher end side of San Jose, just 20 minutes away from the airport, and surrounded by the best gourmet restaurant in the region. Offers mid range 3 star accommodations, trendy bar-restaurant, souvenir shop, adventure tour desk, free delicious continental breakfast and high speed Internet.
• Hotel Santo Tomas, Av. 7 between Calle 3 and 5, Barrio Amon, Tel: (506) 255-0448 Fax: (506) 222-3950, [58]. Pleasant bed and breakfast inn, English speaking staff, single rooms are small, some downstairs rooms quite spacious. Rates from $80, including breakfast.
• Kap's Place, Street 19, Avenues 11 and 13, #1142 in Barrio Aranjuez, Tel +506 221-1169, info@kapsplace.com, [59]. Guesthouse with 13 rooms and 1 large apartment. $30 - $80 including taxes. Guests have free kitchen use and free travel planning help! The K in Kap's Place stands for Karla. It's her house and her service is top notch. Bilingual (English/Spanish).
• Marcela Bell, homestay@racsa.co.cr, arranges accommodation in private homes; will advise on all aspects of visiting Costa Rica.
• Residence Inn San Jose Escazu, tel. 506 2588 4300, [60] 15 minutes from downtown San Jose, complimentary hot breakfast daily, outdoor pool and fully functional gym. The Residence Inn San Jose also has two meeting rooms making it the perfect San Jose hotel for business or pleasure.
• Rincón de San José, (formerly Hotel Edelweiss), Avenida 9 Calle 15 Barrio Otoya, Tel +506 221-9702, info@hotelrincondesanjose.com, [61]. 27 rooms, $50 + tax for a double including breakfast. 10% cash discount. Free Internet.
• TRYP San José Sabana Hotel, Avenue 3, calles 38 y 40, San José COSTA RICA, + (506) 2547 2323; US toll free number 888 4115 487, [62]. Centrally located hotel with conference facilities. (9.9426642,-84.092676) edit
[edit] Splurge
• Barcelo San Jose Palacio, [63]. Located just a short way between San Jose and Heredia, Barcelo San Jose Palacio is just minutes from the International Airport and downtown, and is an ideal starting point for those who fly into the Costa Rica capital to begin their visit to this tropical Central American nation. edit
• Costa Rica Marriott Hotel, (700 Metros east of the Firestone), + 506 2298 0000, [64]. The Costa Rica Marriott is a beautiful colonial hacienda set on a 30 acre Coffee Plantation. Comfortable accommodations, superb restaurants and catering, two outdoor tropical pools, Spa and more. edit
• Doubletree Cariari by Hilton San Jose, Canas Highway San Antonio de Belen (5 minutes from the Airport), (506) 2239 0022, [65]. Distinctive architecture with lots of plants and a great swimming pool $89-$229. edit
• Finca Rosa Blanca, Santa Barbera de Heredia, +(506) 269-9392, [66]. The 13 room Finca Rosa Blanca Country Inn, situated above the beautiful Central Valley of Costa Rica, is surrounded by exotic flora and grows some of the world’s best organic coffee. Finca Rosa Blanca is nestled in 10 acres amidst hundreds of fruit trees and dramatic tropical flora, and offers spectacular views of volcanoes, cloud forests and coffee plantations. The Inn has an a la carte restaurant, Spa and small gift shop on site. $160-$240 for a double. edit
• Hotel Don Carlos, Calle 9 & Avenida 9, Barrio Amón, Tel: 866 675 9259 (Toll Free) or +506 221 6707, [67]. Beautifully decorated hotel, run by the same family for three generations. $70 + 16.39% tax for a standard double. There is also a tour operator (expensive) and a nice souvenir shop inside the hotel.
• Hotel Fleur de Lys, (50m north of Drs. Echandi Clinic), (506) 223-1206, [68]. Comfortable European style small hotel with eclectic class and grace. Excellent restaurant on site, suites feature jacuzzis. edit
• Hotel Grano de Oro, (Just off Paseo Colon), (506) 255-3322, [69]. Converted from a Tropical Victorian mansion, the 35-room hotel maintains the warmth and comfort of a private home. Hallways lined with period photographs and original art meander through the building and present lush tropical flower arrangements and luxuriant plants at every turn. Italian tile fountains grace intimate atriums, ensuring that one is never far from the soothing melody of falling water. The restaurant is beautiful, opening upon a courtyard with a fountain at the center. Service is prompt and efficient, and the staff mostly speak very serviceable, if not near-perfect, English. The rooms are comfortable, but most lack air conditioning which means you will be awakened by bird calls most mornings. Carved woods dominate the public rooms, a beautiful venue which appears to be popular with Ticos as well. edit
• Hotel Villas Corcovado, [70]. Villas Corcovado is a beautiful community of luxurious villas found throughout 70 oceanfront acres of unspoiled rainforest on the Southern Pacific Coast of Costa Rica. Perfect for those who seek adventure, flora and animal life in the inspiring framework of the Golfo Dulce.
• Hampton Inn & Suites, [71]. An ideal accommodation for anyone visiting Costa Rica on business of for those who are set to fly out. Hampton Inn & Suites is located just minutes from the International Airport and is close to many Costa Rican company's headquarters.
• Real InterContinental Hotel and Club Tower Costa Rica, +506 289 7000, [72]. Located amid tropical gardens, only 15 minutes from Juan Santamaria Int’l Airport and 10 minutes from downtown San José, the hotel is close to an array of stores, movie theaters and fast-food restaurants and faces the modern Multiplaza Mall. It has 210 luxurious rooms, 54 rooms of the Camino Real Club, 5 junior suites and a presidential suite, for a total of 261 rooms. Each room is equipped with a mini-bar, hair dryer, air conditioning and a system of electronic key. Services : gym, swimming pool, jewelry shop, tennis court, restaurant, bar, conference rooms and gift shop. edit
• Xandari Resort & Spa, [73]. Xandari Resort and Spa is home to 21 spacious villas on a 40-acre coffee and fruit plantation overlooking the Central Valley. The colorful villas, private terraces, furniture and gardens are designed with great attention to detail, and each villa is adorned with original works of art. edit
• Cristal Ballena Resort and Spa, on the Pacific Coast between Dominical and Palmar Sur. Nature Air and Sansa fly into Palmar Sur, and the Resort can arrange pickup. The owners are an Austrian couple who "retired to go to work," and opened this beautiful resort just above the Pacific ocean with incredible views. Even in the off season, Waldemar will usually be in the restaurant in the evening making sure the guests are happy. Many tours and adventures are available, as usual. The restaurant is very good, one of the better ones in the area. Some Austrian dishes complement local seafood. The view is incredible, every room has a private balcony with a view of the pacific. Monkeys can be heard howling in the morning, and tropical birds are common.
• Quality Hotel Real San José, El Paseo Real, Costado Este Centro Empresarial Forum, Santa Ana., +506 2204 6700 (fax: +506 2204 6800), [74]. Quality Hotel Real San Jose has 154 rooms, 16 mini-suites and rooms for disabled people. Also it has a restaurant, pool, business center with computers, printers and internet access high speed. Prices range between $80 - $140. edit
[edit] Near the Airport
• Adventure Inn, Tel: +506-2239-2633, [75]. Only 8 minutes from the airport with free transportation. $77 single, $94 double, includes an all you can eat American breakfast, free airport transportation, free international phone calls, on-site sports bar / restaurant, pool and Jacuzzi, fully-equipped gym, hotel-wide wireless internet. Click here for a 10% discount off your room rates for cash or travelers cheques or mention WikiTravel at your time of booking.
• Hotel Aeropuerto, 50624337333, [76]. checkin: 13.00; checkout: 12.00. An Airport hotel surrounded by nature, more than 40 property extension is covered by trees and gardens. Hotel Aeropuerto is located near the international airport san jose costa rica With pool, restaurant, tours, room service, free internet (wifi), currency exchange, very secure facilities, free airport shuttle 24/7 . 80. (95,) edit
• Hampton Inn. $105/night avg (from Travelocity) Right in front of the airport, adjacent to Fiesta Casino and a Dennys restaurant.
• Trapp Family Country Inn, 786-8379198, [77]. checkin: 13.00; checkout: 12.00. The Trapp Family Country Inn is located only 1 mile from Juan Santamaría International Airport. Comfortable accommodations along with genuine family service in a peaceful atmosphere. This colonial style hotel is surrounded by beautiful gardens with centenary fig trees. 80. (95,) edit
• Vina Romantica Bed and Breakfast, (506) 2430-7621, [78]. Great entry/exit point for travelers, minutes away from main tourist pick up points and the airport. Provides comfortable rooms with memory foam mattresses, private bathrooms, restaurant and beautiful surroundings such as Doka Coffee fields and Poas Volcano. edit
[edit] Stay safe
The traffic is dangerous. Cars don't stop for pedestrians, and they generally drive very fast. The area around the Coca Cola Bus Terminal is not safe either during the day or at night. You should watch your belongings and stay with a group at all times in the city.
Most Ticos (Costa Rican natives) are friendly and honest. However, if you have a flat tire on the main highway to the airport, don't accept help except from an established service station, and never accept help from a person who offers to help on the highway. It may become a car-jacking. Petty theft is high risk, including from valet parking staff and housekeepers in hotels. Carry a cell phone and know the number of the local "policia".
If you travel by bus, never(!) put your luggage into the storage space, even if the driver wants you to at the beginning of the travel. The bus usually stops every 5 minutes and picks up people from the street. But there are also people who open the space while you are up in the bus, pull out a rucksack (backpack) and disappear. If you put your rucksack between your knees, nothing will happen. Car theft is common so if you drive (as you can) from the US, or wherever else, bring a club (steering wheel lock) or park in locked fence areas. Most houses inside San Jose have bars on the windows and large gates in which to put vehicles. Despite the numerous pot holes and other bumps in the street, the drivers have a tendency to speed while the luggage and passengers are getting thrown around so please wear your seat belt just in case. Going off of the scenarios of dangerous drivers, the taxi cabs, as well as any vehicle for that matter, that you choose to travel in MUST have a yellow triangle sticker on the front doors with the plate number. This demonstrates that that particular vehicle and the driver are legitimate. If anything were to happen you (as a tourist) would know who to file a complaint etc with. Just be advised: Please do not enter any vehicle that does not have this larger triangle sticker on it (usually on the door) because the driver is most likely an illegal taxi or someone trying to con you. The airport is a frequent "hot-spot" for these sort of incidents to occur quickly, because someone can come up to you (knowing that you are a tourist, not aware of their customs or regulations, and will exploit that), grab your luggage, and start loading it into their car - but be sure to check the vehicle for the yellow triangle as well as wrestling your luggage back out of the car. The orange taxis at the airport are the official airport taxis.
Crime is a serious problem so be constantly alert and aware of your surroundings and use your common sense. Be particularly careful with your passport. Also, police may stop you and take you to jail for not carrying your passport, or a photocopy of the main page and the entry stamp (very reccomendable instead of carrying the passport with you). A tour bus was stopped in Limon in January 2006 and several US citizens were taken to jail for not having their passports on them, but this is not common.
San Jose, as the largest city in C.R. has the largest hospitals, both public and private. Tourists can use the private hospitals, and pay with cash or credit card. The wait is significantly shorter than at public hospitals. Also note the bigger private hospitals in the country are considerably more expensive than the many, smaller private hospitals throughout the city. Most doctors can speak medical English, and they provide translator services. Most private doctors and hospitals do take foreign insurance plans. Ask before. If you are unlucky enough to have your child get really sick he or she will be transferred to the only children's hospital in the country in San Jose, which is public.
Generally speaking if you stick to the tourist spots in the city you will be safe, just try to avoid showing off valuables more than necessary, if you're taking a picture put your camera away as soon as its taken, never show big amounts of cash, exercise caution. Avoid at all costs walking at night, either right downtown or in the suburbs, cabs and buses are too cheap, so walking at night is a very unnecesary risk. As with any big city, use common sense and keep your belongings in front or beside you - never on your back. San Jose is known for its abundance and skill of pickpocketers.
[edit] Contact
[edit] TV
Cable TV channels have many American English language channels includings Fox News, CNN, CNBC, TNT, HBO, ESPN, ABC, NBC, and CBS.
On Amnet in San Jose ABC, CBS, and NBC are broadcast on channels 69-71, respectively.
[edit] Cope
[edit] Embassies and High Commissions
• China, (De la casa de D. Oscar Arias 100 metros al sur y 50 metros al oeste, Rohrmoser, Pavas San Jose), +506 22914811 (fax: +506 22914820), [80]. edit
• Japan, Torre la Sabana Piso 10 (Sabana Norte, 300 m oeste y 25 m norte del I.C.E.), +506 2232-1255 (fax: +506 2231-3140), [81]. edit
• United States, Calle 120 Avenida 0, Pavas, +506 2519-2000 (fax: +506 2519-2305), [82]. edit
[edit] Get out
Being the capital, San José is the hub for all travel in Costa Rica. You can take the local buses or the more expensive bus services from Interbus or Gray Line Fantasy Bus. There is also a lot of smaller airports dotted around the country for minor destination hopping.
There is an exit tax (US$26 for both visitors and residents) to leave the country. You can pay for it at a bank (any BCR or Banco Nacional) and get a receipt or wait in the line at the airport. Pay the tax as soon as you arrive at the airport so you don't miss your flight and arrive with cash. Be sure to have a pen, as there is a small form to fill that is going to be requested before check-in. Tobías Bolaños Airport in Pavas serves as Nature Air's hub for flights within the country and also has 4 flights per week to Bocas del Toro, Panama. Nature Air used to fly to Granada, Nicaragua as well, but as of April 2007, the flights have been discontinued due to the poor condition of the runway in Granada.
AirPanama [83] flies from Juan Santamaría airport to David/Panama three times a week.
A trip to Jacó, on the Pacific Coast, is a delight, although the highway that goes there is not a superhighway. Surfing is great, and since many young chefs are surfers, you can get some really great dining at amazingly low prices. A little farther down the coast, a stop at Manuel Antonio National Park is a must.
On your way to Jacó, stop for lunch at "Mirador del Cafetal" (View of the Coffee Plantation), just beyond Atenas. Views are spectacular!
• About 3.5-hours by car or 20 minutes by plane south of San Jose lies Manuel Antonio National Park in Quepos, Puntarenas. Of Costa Rica's dozens of national parks, Manuel Antonio has long been one of the jewels, an idyllic combination of exuberant forest, white-sandy beaches, and rich coral reefs. The guardians of this beautiful wilderness are now attempting to harness its popularity by limiting the number of ecotourists. This park is one of the country's smallest and only remaining habitats for the red-backed squirrel monkey. Manuel Antonio is located along the coast line and offers snorkeling, skin diving, surfing, and fishing galore. After a visit to the rain forest that gets you hot and sweaty, nothing beats jumping in to the refreshing ocean. Open Tu-Su.
• An easy 2-hour drive north-west of the capital, Costa Rica's most favored rain forest getaways in La Fortuna, San Carlos. The Chachagua Rain Forest Hotel is in the Tilarán mountain range. One of the most biologically diverse in the entire country, the Chachagua's 50-acre spread nestles up against the Children's International Rain Forest which has a great potential for bird and wildlife viewing. The Arenal Volcano and Lake are within driving distance. There is spelunking, white-water rafting, and rappeling for those who aspire to do it all, and the Tabacón Hot Springs for those who do not. Chachagua is fairly self-sustaining: it grows its own fruit and other produce is grown on the ranch. Overall the hotel provides nature guides, nature tours, walks, breakfast, dinner, and a transfer to/ or from San Jose. The best times to visit are November through March. Call Chachagua Rain Forest Hotel at (Tel) 506/239-6464 or fax 506/293-8553 for more information.
This is a usable article. It has information for getting in as well as some complete entries for restaurants and hotels. An adventurous person could use this article, but please plunge forward and help it grow!
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Difference between revisions of "Los Angeles/Downtown"
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(updated listing Yang Chow)
(added listing Rodeway Inn and Suites Convention Center Los Angeles)
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Downtown has a marvelous selection of excellent hotels that cater primarily to business travelers. That means that if you time your visit right (for example use weekends), you can get a great room at a very reasonable price.
Downtown has a marvelous selection of excellent hotels that cater primarily to business travelers. That means that if you time your visit right (for example use weekends), you can get a great room at a very reasonable price.
+
+
*<sleep name="Rodeway Inn and Suites Convention Center Los Angeles" alt="" address="1904 W. Olympic Blvd." directions="" phone="213-380-9393" url="http://www.laconventioninn.com/" checkin="" checkout="" price="" lat="" long="">Welcome to Rodeway Inn Convention Center Los Angeles. Our reasonably priced motel is ideal for a variety of travelers, and you'll find ample entertainment and attractions in and around our downtown neighborhood.</sleep>
=== Budget ===
=== Budget ===
Revision as of 18:28, 17 September 2012
Los Angeles City Hall
L.A. is a city of diverse cultures and many are showcased in or around Downtown. The area's highlights include Grand Central Market, Museum of Contemporary Art, Disney Concert Hall, The Music Center, Olvera Street, Chinatown, Little Tokyo, and the Japanese-American Museum. Downtown is also home to some of the most unique and stunning examples of American and international architecture.
Contents
Get in
Downtown LA is simultaneously the hub of the freeway network, road network, commuter rail network, subway / light rail network, and bus network in the region. It is thus accessible from multiple entry points.
By freeway
Downtown LA can be accessed directly via the Pasadena Freeway (SR-110), the Santa Monica Freeway (I-10), and the Santa Ana Freeway (I-5 and US-101). Just outside Downtown LA, these freeways connect to the Golden State Freeway (I-5), the Hollywood Freeway (US-101), the San Bernardino Freeway (I-10), the Harbor Freeway (I-110), and the Pomona Freeway (SR-60).
Drive your car to a parking lot and go by foot from then on. Downtown isn't that big. Most likely, a DASH shuttle has a stop where you want to go.
By commuter rail
If your point of origin is within the urban and suburban areas of Los Angeles, Ventura, San Bernardino, Riverside, Orange, or San Diego Counties, you may be able to avail of the growing commuter rail network known as Metrolink [1] to visit Downtown LA. Six of the seven Metrolink commuter rail lines terminate at Union Station in Downtown LA's El Pueblo district. Tickets can be purchased from vending machines at each station, and fares are determined by time (peak or non-peak hour, weekday or weekend) and distance:
• The San Bernardino Line runs 34 trains on weekdays between Downtown LA and the eastern suburbs (the "Inland Empire"), running through the San Gabriel and Pomona Valleys, and San Bernardino County. There is also limited weekend service, and terminating in Downtown San Bernardino.
• The Riverside Line runs 12 trains on weekdays between Downtown LA and the eastern suburbs (the "Inland Empire"), running through the San Gabriel and Pomona Valleys, and Riverside County, and terminating in Downtown Riverside.
• The 91 Line runs 8 trains on weekdays between Downtown LA and the eastern and southeastern suburbs (the "Inland Empire"), running through the Orange and Riverside Counties, and terminating in Downtown Riverside.
• The Orange County Line runs 20 trains on weekdays between Downtown LA and the southeastern suburbs, running through Orange and San Diego Counties, and terminating in Downtown Oceanside. There is also limited weekend service.
• The Ventura County Line runs 20 trains on weekdays between Downtown LA and the northwestern suburbs, running through the San Fernando Valley and Ventura County, and terminating in Ventura's Montalvo neighborhood.
• The Antelope Valley Line runs 24 trains on weekdays between Downtown LA and the northern suburbs, running through the San Fernando, Santa Clarita, and Antelope Valleys, and terminating in Downtown Lancaster. There is also limited Saturday service.
By subway / light rail
For those visitors coming from within Los Angeles County, local subway and light rail service may be the best option to get to Downtown LA. Four of the five subway and light rail lines in the Metro Rail [2] system terminate in Downtown LA's Union Station and 7th/Metro Center. Each boarding requires a $1.50 fare. For unlimited rides on Metro Bus and Metro Rail, purchase a $6 day pass on any Metro Bus or in any Metro Rail station.
• The Red Line subway brings riders from the San Fernando Valley, Hollywood, and parts of the Eastside, cutting across the Financial District, the Jewelry District, the Civic Center, and El Pueblo, passing near the Historic Core and Bunker Hill, making four stops in Downtown LA (7th/Metro Center, Pershing Square, Civic Center, and Union Station).
• The Purple Line subway brings riders from Mid-Wilshire, joining with the Red Line subway to cut across the Financial District, the Jewelry District, the Civic Center, and El Pueblo, passing near the Historic Core and Bunker Hill, making four stops in Downtown LA (7th/Metro Center, Pershing Square, Civic Center, and Union Station).
• The Blue Line light rail brings riders from South Los Angeles (also known as "South Central"), Long Beach, and other southern suburbs, to two Downtown LA stops: Pico Station in the South Park district, and 7th/Metro Center in the Financial District.
• The Gold Line light rail brings riders from Pasadena, Boyle Heights and East Los Angeles to three Downtown LA stops: Chinatown Station in the Old Chinatown district just north of the Civic Center, Union Station, and Little Tokyo/Arts District Station in Little Tokyo.
• There is also a Green Line connecting the city of Norwalk to the beach area of Redondo/Hermosa with connections to the Blue Line and the LAX shuttle bus.
By bus
By air
Downtown LA is not directly served by an airport, but can be accessed via public transportation from Los Angeles International Airport (IATA: LAX) in Westchester and Bob Hope Airport in Burbank.
From LAX in Westchester
From LAX Airport [3] there are two relatively frequent public transportation options to Downtown LA: the FlyAway Bus, and Metro Rail.
• For the FlyAway Bus option, simply walk to the nearest platform with a green shuttle sign outside your terminal. The FlyAway Bus will stop at each terminal to pick up passengers bound for Van Nuys, Westwood, and Union Station. Be sure to board the Union Station-bound FlyAway Bus, or you will end up far away from Downtown LA. This express bus uses the carpool lanes and busways on major freeways from LAX straight downtown, and costs $7 each way. Buses run 24 hours a day, 7 days a week.
• For the Metro Rail option, which is usually slower due to the many transfers necessary, first board a free LAX G-shuttle to the Metro Green Line platform at Aviation Station. Then take a Norwalk-bound Green Line train to Imperial/Wilmington station, where you will walk down the stairs and board a Los Angeles-bound Blue Line train. The Blue Line ends in the Financial District, at 7th/Metro Center, connecting to the Red and Purple Lines. Each boarding costs $1.25. If you are planning on using the system further, it may be worth it to purchase a $5 Day Pass before boarding the Green Line train. Trains run until about 1AM everyday.
From Bob Hope Airport in Burbank
Bob Hope Airport is served by two rail networks, both of which use the Burbank Airport train station - walking distance from the main terminal building.
• Metrolink commuter rail: Use the ticket vending machines to purchase a ticket from Burbank-Bob Hope Airport to Union Station, the main train station downtown. Metrolink runs 15 trains in each direction on the Ventura County Line on weekdays during commute times (use Amtrak at night and on weekends). See the Metrolink Burbank-Bob Hope Airport schedules on the web [4] for exact departure times. This trip costs $5.25 each way and takes from 14 to 31 minutes.
• Amtrak: Use the ticket vending machines to purchase a ticket from Burbank-Bob Hope Airport to Union Station, the main train station downtown. Amtrak runs 5 trains in each direction on the Pacific Surfliner Line 7 days a week, with the last train departing at 9:13PM from Burbank-Bob Hope Airport. This trip costs $4 and takes about 26-43 minutes.
If you must drive, park at the NE corner of Hill and 9th to check out the Fashion/Garment District. Incredible deals, great restaurants, beautiful architecture (check out the Orpheum and Eastern Columbia Buildings across from the lot). There is also Cliftons further up Broadway, The Arcade Building which, like many of the historic building downtown, is being converted into upscale lofts.
• Parking: Some people are partial to parking at any one of the lots around the Music Center or Civic Center in roughly the area bounded by Grand to Spring and Temple to 2nd. But Pershing Square has good centralized parking. If you are checking out the Convention Center (only do if you are actually going to a convention there) consider parking there, although it is adjacent to Staples Center, which is a block from the Hotel Figueroa (check out the bar and the Moorish architecture), which is a block from the Pantry...well, you get the idea.
Get around
Downtown is probably the only part of L.A. that one can reasonably cover on foot.
• Metro Bus[5] is the most extensive bus system in the region. All major streets have at least one (and in some cases, several) bus lines running daily. Base fare is $1.50 and an unlimited-use day pass costs $5. Both can be purchased on board any Metro bus.
• Metro Rail[6] is the subway and light rail system for Los Angeles County. Downtown LA can be traversed using the six downtown rail stations that are served by four of the five lines in the Metro Rail system. At the northern end of Downtown LA, the Gold Line stops at Chinatown on its way northeast to Pasadena. The Red and Purple Line subways meet with the Gold Line light rail in Union Station, where connections can be made to buses, Metrolink commuter trains, and Amtrak. From Union Station, the Red and Purple Line subways run along Hill Street, making stops at the Civic Center and Pershing Square, before turning west under the Financial District. There they connect to the Blue Line light rail at 7th/Metro Center. From there the Red and Purple Lines run northwest and west, respectively, and the Blue Line runs south through Downtown LA's redeveloping South Park district, with a stop at Pico, towards the city of Long Beach.
• DASH[7] is a shuttle service run by L.A. Department of Transportation. When your feet get tired or to better expand your travel area use the DASH. It has several convenient routes that whisk you to almost all of the worthwhile spots Downtown. A ride currently costs 50 cents (25 cents for seniors) and pamphlets are available from most MTA stations (Union, 7th/Olive) and convenience stores Downtown (spotty weekend and after-hour service though).
See
Cultural districts
Olvera Street
• Chinatown, [8]. Primarily centered around North Broadway; unlike Chinatowns in many other cities, it has a wide, main, busy street filled with small shops and restaurants. At about the middle point of N. Broadway in Chinatown is an open market much like those found in Hong Kong. Be sure to haggle!
• Little Tokyo, [9]. Also known as J-Town, the Japanese district features restaurants, museums, and shops. It sits in the area between Temple and about 5th and Spring through Alameda.
• Olvera Street, [10]. This is where LA was founded as El Pueblo de Los Angeles. You can take a tour of the city's oldest house to see what it looked like at that time. The plaza is mostly filled with Mexican trinket stands and Mexican restaurants.
Museums
• Museum of Contemporary Art (MOCA), 250 S. Grand Avenue, Los Angeles, CA, 90012, +1-213-626-6222, [11]. Th-M: 11AM-5PM. The permanent collection is fairly interesting, but the changing exhibitions can be more hit or miss. The museum has no 'traditional' art, so bring an open mind. The gift shop (free entrance) is fun for at least 20 minutes of wonder and awe. $12, $7 student (includes admission to Geffen Contemporary).
• Geffen Contemporary, 152 N. Central Avenue, Los Angeles, CA, 90013, [12]. A branch of MOCA tucked away in Little Tokyo. Same opening hours and shared tickets as MOCA on Grand.
• Japanese American National Museum, 369 E. First Street, Los Angeles, CA, 90012, +1-213-625-0414, [13]. Tu-Su: 10AM-5PM. Covers the Japanese-American experience, with a special emphasis on the concentration camps of World War II. $8.
• Old Plaza Firehouse, 134 Paseo de la Plaza, +1 213 625-3741. Tu-F: 10AM-3PM, Sa-Su: 10AM-4:30PM. This was the original fire station for the City of Los Angeles. Built in 1884, it has been restored to its original condition. The knowledgeable docents offer a peek into Los Angeles in the 19th Century. Free (donations accepted).
• Grammy Museum, 800 W. Olympic Boulevard (entrance on Figueroa St), +1-213-765-6800, [14]. M-F 11:30AM-7:30PM, Sa-Su 10AM-7:30PM. History of music, with listening posts. Adult $12.95.
Arts
Walt Disney Concert Hall
• Fashion Institute of Design & Merchandising (FIDM), 919 S. Grand Avenue, Los Angeles, CA, 90015, +1-800-624-1200, +1-213-624-1201, [15]. Gorgeous campus of FIDM and ongoing free exhibits make this a pleasant way to kill a couple of hours.
• The Los Angeles Central Public Library, 630 W. 5th Street, Los Angeles, CA, 90071, +1-213-228-7000, [16]. Huge library rebuilt in the 1980s and '90s. Almost always has a public exhibition going.
• Music Center and Disney Hall, 135 N. Grand Avenue, Los Angeles, CA, 90012, +1-213-972-7211 (), [17]. Impressive hall architecture complete with tours most days. The Dorothy Chandler Pavilion is open to the public Christmas Eve day with almost round the clock performances by amateur cultural arts groups. The Walt Disney Hall has daily tours, check website for schedules.
Architecture
• The Bradbury Building, 304 South Broadway, Los Angeles, CA, 90013. Built in 1893, the Bradbury Building is one of Southern California's most remarkable architectural achievements. Behind its modest exterior lies a magical light-filled Victorian court that rises 50 feet with open cage elevators, marble stairs and ornate iron railings. The building has been a set for many movies, including Blade Runner in 1982. Visitors without business in the building are allowed into the lobby and up to the first landing of the staircase.
Cathedral of Our Lady of Angels
• The Cathedral of Our Lady of the Angels, 555 W. Temple Street (between Grand Ave & Hill St), +1 213 680-5200 (), [18]. 6:30AM-6PM M-F, 9AM-6PM, Sa 7AM-6PM Su, hours extended to 7PM during daylight savings time. This large and austere cathedral, dedicated to Saint Vibiana, is the head of the Archdiocese of Los Angeles. It was opened in 2002 at a cost of nearly $200 million, replacing The Cathedral of St Vibiana which was heavily damaged in the 1994 earthquake.
• Library Tower (US Bank Tower), 633 W. Fifth Street, Los Angeles, CA, 90071 (across Fifth Street from the downtown central library). At 73 floors and 1,017 feet, it is said to be the tallest building between Chicago and Hong Kong. Note to photographers: the Library Tower's security personnel will try to discourage you from taking pictures of this building. As long as you are standing on a public sidewalk you may legally take any picture you like in the United States.
• St. Vincent court, 7th Street, between Broadway and Hill, [19]. A tranquil hideaway tucked in the heart of the Jewelry District.
• The Theater District. The Theater District along Broadway has been converted to discount jewelry, electronics and ethnic shops, but much of the architecture and the marquees remain.
• Union Station. No trip to downtown LA would be complete without a visit to the historic train station, built in 1939 with a Spanish mission exterior. The large waiting room and restaurant is like it was in the 1940s. It is used in lots of movies, including Blade Runner, where the main hall was used as the police station.
Do
• Arroyo Seco Historic Parkway (110 Freeway), (starts at the intersection of the CA-110 and the CA-101, heading north from that junction), [20]. Drive the Parkway, a National Scenic Byway that runs for 9.4 miles (15.1 km) between Downtown Los Angeles and Pasadena. The Parkway passes from the skyscrapers of Downtown, through Chinatown into the Arts-and-Crafts style neighborhoods of South Pasadena and ends in Pasadena at Colorado Blvd., home to the famous Rose Parade.
Tours
Various groups offer free or cheap walking tours of Downtown LA.
• Downtown Art Walk, [21]. A free monthly self guided tour--and free walking tours, reservations required--held on the second Thursday of every month, to art galleries and museums in Downtown L.A.
• L.A. Conservancy Walking Tours, [22]. See the grand Vaudeville/Movie theaters of the 20s and the impressive Art Deco office buildings in several easy to handle walking tours. Strongly recommended for those wanting to grasp a feel of LA's history. Reservations are strongly recommended.
• Las Angelitas del Pueblo, [23]. This is a group of volunteer docents who give free tours of El Pueblo de Los Angeles to the public.
• Raymond Chandler's Los Angeles: In A Lonely Place bus tour, [24]. An occasional bus tour of sites downtown and in Hollywood from the films, books and lives of Raymond Chandler and his anti-hero Philip Marlowe. $58, includes snacks.
• The Real Black Dahlia bus tour, [25]. A true crime and social history tour that intimately explores the last weeks of Elizabeth Short's life, asking not "who killed her?" but "who was she?" $58.
• John Fante's Dreams of Bunker Hill bus and walking tour, [26]. An occasional bus and walking tour of sites downtown and in Hollywood from the life and work of novelist John Fante and his great fan Charles Bukowski, plus crime scenes from forgotten horrors of old Bunker Hill, Sonora Town and beyond. $58, includes snacks.
Sports
• Staples Center, [27]. Home to five of LA's pro sports franchises; Lakers (NBA), Kings (NHL), Clippers (NBA), Avengers (AFL), and Sparks (WNBA), plus many concerts, shows and conventions.
• Dodger Stadium - Home to the Los Angeles Dodgers, Major League Baseball franchise of the National League.
• Los Angeles Memorial Coliseum - Home to the University of Southern California Trojans NCAA football team, members of the Pacific Twelve conference. Site was host to the Olympics in 1932 and 1984, the home to LA's former NFL franchises the Los Angeles Rams and Raiders from 1946-1979 and 1982-1994 respectively, and the home of UCLA football until 1982, when they moved to the Rose Bowl.
Buy
Downtown's shopping districts are sights in themselves.
• Fashion District,[28] Where style and cheap textiles smash together. Important for the addicted shopper. You can find the district in the Southeast corner of Downtown roughly where Spring and Main meet going Southeast.
• Flower District (766 Wall Street} [29], The best place to get the best cut and potted flowers and plants, plus just a great site to see.
• Jewelry District,[30] Wonder where all of those West Coast Rappers get their bling bling? Well, if they are frugal, they get it in the Jewelry District. Bounded by Olive-Broadway and 6th-7th, it is conveniently close to Pershing Square (parking and Red line access).
Some shops stand out:
• Mikawaya (800 E. 4th St. Little Tokyo) [31], Their moto says it best: "The finest name in Japanese pastries since 1910"
• Capucci Optics (7th+Fig Mall), Great place to get a pair of great glasses, sunglasses or contacts at a reasonable price. Ask for Fatima for friendly service.
• Santee Alley (Fashion District), Home of knock off designer labels and everything else you could possibly imagine, located between Santee Street and Maple Avenue, starting on Olympic Boulevard.
Eat
This guide uses the following price ranges for a typical meal for one, including soft drink:
Budget Under $10
Mid-range $10 - $25
Splurge Over $25
Budget
• Clifton's Cafeteria, 648 S. Broadway, Los Angeles, CA, 90014 (Downtown), +1-213-627-1673, [32]. Daily: 6:30AM-7:30PM. Since 1935, located on Broadway, serves cafeteria style food. One should experience the history, the food at affordable prices, and of course view the redwood forest theme.
• Cole's Pacific Electric Buffet, 118 E. 6th Street, Los Angeles, CA, 90014 (on 6th, between Main and Los Angeles), +1-213-622-4090 (), [33]. Daily: 9AM-10PM. Bar/restaurant in nearly continuous operation since 1908, but recently shut for a year and a extensive upscale redesign. Along with Philippe The Original, one of the possible originators of the French Dip sandwich.
• Empress Pavilion, 988 N. Hill Street, Los Angeles, CA, 90012 (Chinatown), +1-213-617-9898. Most people come here for the dim sum on carts but there is also a menu.
• Frying Fish, 120 Japanese Village Plaza (Little Tokyo), 213-680-0567. Sushi restaurant with a food conveyor belt built into the bar, located in the Japanese village. Don't miss the excellent California roll!
• Grand Central Market, 317 S. Broadway, Los Angeles, CA, 90013, +1-213-624-2378, [34]. Daily 9AM-6PM. Huge indoor bazaar of Central and South American vendors. Get fresh tortillas, huge Mexican papayas and tasty Tortas. On Hill and Broadway between 3rd and 4th (closer to 3rd). Conveniently near the Bradbury Building (unique architecture) and the Pershing Square Red line stop (Northeast access).
• The Original Pantry Cafe, 877 S. Figueroa Street, Los Angeles, CA, 90017, +1-213-972-9279. The Pantry boasts that it has never closed or been without a customer since it first opened in 1924. (Want proof? The front entrance has no lock on it). Come here on any morning and you will see a line stretching around the block - the wait is worth it, and the fast service will have hot plate of food in front of you within minutes of sitting down. Best place for breakfast after midnight. Cash only.
• Original Tommy's, 2575 W. Beverly Blvd, Los Angeles, CA, 90057 (On the corner of Beverly and Rampart just west of Downtown Los Angeles), +1-213-389-9060, [35]. Open 24 hours/7 days a week. A Los Angeles landmark since 1946 Tommy's is a can't-miss for any hamburger lover. Serving hamburgers, french fries, hot dogs, and tamales with their "secret blend" of chili you will always find a line for food at all hours, especially late night/early mornings.
Philippe's
• Philippe's, 1001 N. Alameda Street, Los Angeles, CA, 90012 (Chinatown, one block from Union Station), +1-213-628-3781, [36]. Daily: 6AM-10PM. Aaaah...an LA landmark situated a couple of blocks north of Olvera St. and Union Station is a nostalgic shop with hay and sawdust covered floors. Famous for their 'French Dip' sandwiches dipped in au jus ($4.90), but the real reason to go is the atmosphere and the pastrami — the joint opened in 1908 and the menu still features things like pickled eggs and pig's feet. Coffee is ten cents a cup, but their 60-cent lemonade is even more popular. Expect to queue at any time and the place is mobbed on the nights of Lakers and Dodgers games.
• Señor Fish, 422 E 1st Street, Los Angeles, CA, 90012, +1-213-625-0566. Not really authentic -- it's sort of a variation on Baja-style Mexican -- Senor Fish downtown does just one thing well, but they do it better than anyone. Luckily, that one thing is an important thing: grilled fish tacos. Grilled, not fried. Their Shrimp Taco is amazing as well.
• Spring Street Smokehouse, 640 N. Spring Street, Los Angeles, CA 90012 (in China Town, on Cesar Chavez and North Spring), +1-213-626-0535, [37]. M-Tu: 10:30AM-8PM, Wednesday-Friday: 10:30AM-9PM, Sat: 12PM-9PM. The best barbecue in town. 27 microbrews.
• Weiland Brewery, 400 E. 1st Street, Los Angeles, CA, 90012 (in Little Tokyo, on Central and First), +1-213-680-2881, [38]. M-F: 11AM-2AM, Sat: 5PM-2AM. The cheese fries are to die for. Very affordable place for drinks. One of the few bars with a weekend happy hour lasting until 2AM
Mid-range
• Engine Co. No. 28, (Figueroa Corridor), [39]. Comfort food at its best. A restored actual fire station that churns out LA's best meatloaf, fried chicken and lemonade, all in an elegant atmosphere with great service.
• J Restaurant & Lounge, 1119 S. Olive St (at 11th St), +1-800-850-6074, [40]. Dining and entertainment, located near the Staples Center. The vibe is equal parts hip and casual; the large space has a glitzy lounge, featuring live music, and a mega patio with fire pit and skyline views of the city. Inspired décor. Menu is Mediterranean-meets-American.
• Kendall's Brasserie, 135 N. Grand Ave (at the Music Center), +1-213-972-7322, [41]. Great French menu at a perfect location to catch any of the great evening programs at the surrounding venues. Whatever you order, do not miss their French Fries! Mains from $15.
• Riordan's Tavern, 875 South Figueroa St., +1 213-627-6879 (), [42]. Good (but slightly pricey) pub food in the heart of downtown near the Staples Center. The Mayor's Burger is a one pound beast with chili, bacon, and all the fixings, or you can try the daily carvery sandwich. Steaks and seafood are also decent, and the drinks are poured stiff. $15-$30.
• Royale, 2619 Wilshire Blvd (inside Wilshire Royale Hotel), +1-213-388-8488. Located in the renovated Wilshire Royale Hotel, Chef Eric Ernest's new, culinary digs features a groovy cocktail lounge and menu that’s described as "sophisticated yet approachable."
• The Wood Spoon, 107 W 9th St, +1-213-629-1765, [43]. M 11AM-3PM, T-Fr 11AM-3PM, 6PM-9PM, Sa 12PM-3PM, 6PM-10PM, closed Sunday. Located in a relatively non-descript setting downtown, this restaurant features Brazilian-inspired dishes that are different from what most American restaurants serve as "Brazilian". Rice, beans and plantains are in use, but entrees such as a Brazilian-inspired pot pie and cinnamon water will be new to most diners. Jacqueline, the very gracious chef, will usually make the rounds once the kitchen closes and can tell some very interesting stories about her life after coming to the States. $10-$20 per person.
• Yang Chow, 819 N Broadway (at Alpine Street), +1-213-625-0811, [44]. Located in Chinatown. Award-winning restaurant. Be sure to order the slippery shrimp and the dry sauteed vegetables (green beans and asparagus).
• Yorkshire Grill, 610 W 6th St (6th st & Grand), 213-623-3362. M-F 6:00 AM -3:30 PM, SAT 8:00 AM - 2:30 PM. Yorkshire Grill has been operating since 1954, with many a lucrative business deal having been negotiated over the famous Yorkshire pastrami sandwich. Open early, the Yorkshire breakfast dishes are some of the best in the area and their old school diner coffee will get you off to a strong start to your day! Lunch is always packed at Yorkshire so be sure to get there early, however Yorkshire also offers delivery to your home or place of business. $10.
• Zucca, 801 S. Figueroa St (at Eighth Ave), +1-213-614-7800. Joachim Splichal (of Patina) and chef Giancarlo Gottardo strike the right chord with their sleek, alluring bistro featuring classic Italian fare. The pastas and fresh fish are wonderful - one entrée representing every major region in Italy. Between the cuisine and pleasing milieu, it's quite a lovely dining experience.
Splurge
• Cafe Pinot, (Central Library Courtyard), [45]. A romantic French/Italian restaurant and a unique setting as part of the central library's front yard.
• Ciao Trattoria, 815 W. Seventh St (near Figueroa), [46]. Harry Hagani's homage to fantastic Italian food is a cozy and elegant restaurant popular at lunchtime with the busy executive crowd.
• Cicada, 617 S. Olive St (at 7th St), +1-213-488-9488. M-Fr 5:30PM-9PM. Situated in the beautiful Arts Deco Oviatt Building, Cicada deftly blends elegance of design and superior Italian fare. A chic bar is upstairs, complete with marble dance floor. A perfect place for special occasions, a fine meal before the theatre or just any excuse to be dazzled, both by the atmosphere and the cooking.
• Nick and Stef's, 330 South Hope St, [47]. Fantastic steak house, run by the Patina restaurant empire. If you like beef, this is some of the best in town, with a glass-enclosed aging room where you can view the meat as it ages. Try the dry-aged Ribeye, it will make your head spin. They also have 12 kinds of potatoes on the menu. Not sure why, but they're all good. In the Wells Fargo Center, across from MOCA.
• Pacific Dining Car, 1310 West 6th St, [48]. Don't be surprised if you run into a city politician or other public figuers in this LA landmark that is located partly inside a railway train car, and has been open since 1921. Ask for the breakfast menu any time, day or night, for a more affordable and quite delicious menu.
• The Palm, (across from the Staples Center), [49]. The Palm is a casual white tablecloth restaurant with a mix of Italian, seafood and great steaks. Check out the collection of caricatures on the walls too.
• Traxx, (Union Station). Fancy-Schmansy restaurant in Union Station. Good food, pricey but the ambiance of Union Station makes it worth a splurge.
• Water Grill, (the Old Bank District), [50]. The best seafood and overall service period. Perhaps a bit pricey, but elegant and wonderful.
Drink
Cafés
Alcohol
• Broadway Bar, [51]. Dark, popular spot featuring a bar in-the-round and a second floor smoking balcony.
• Ciudad, [52]. Great mojitos and live jazz music on certain nights.
• Club Mayan, [53]. Best singles spot downtown! Dress code is enforced. Be sure to check out the annual Lucha Vavoom (lucha libre and burlesque).
• Elevate Lounge @ Penthouse, 811 Wilshire Blvd, +1-213-623-7100, [54]. Excellent views of the city.
• Gallery Bar, (inside the Millennium Biltmore Hotel). Upscale cocktail lounge. House drink is the Black Dahlia cocktail, named for the famed victim of the notorious murder, who was last seen wandering through the hotel.
• Golden Gopher. Ms. PacMan + Jukebox + Classiness. Also sells alcohol to-go.
• Grand Avenue Sports Bar, (inside Millennium Biltmore Hotel), [55]. Downtown's best sports bar happens to be located inside its most glamorous hotel. But don't shy away expecting a stuffy joint filled with tuxedo wearing guests from the next door ball, you can relax here in your jeans and enjoy the multiple big screen TV's tuned to your favorite game.
• La Cita. Curious mix of Latinos and hipsters.
• Library Bar, 630 W. 6th Street, +1 213 488-1931. An upscale pub style bar with a floor-to-ceiling and wall-to-wall library that houses a wide range of literary classics, while playing great music from Jim Hendrix to The Who. Knowledgeable bartenders offer an extensive beer selection and cocktails that are both classic and innovative.
• The Rooftop Bar @ The Standard, 550 S. Flower St (at Sixth), [56]. Open daily 12:00PM until 1:30 AM. This unique bar offers a hipster hangout with excellent views of the city from thirteen stories up. Wear warm clothes during cold weather, and be prepared for drink prices in the $10+ range for mixed drinks. Don't forget to try the waterbeds or even jump in their pool for a swim.
• Seven Grand. Popular whiskey bar owned by the owner of the Golden Gopher and Broadway Bar.
Sleep
Downtown has a marvelous selection of excellent hotels that cater primarily to business travelers. That means that if you time your visit right (for example use weekends), you can get a great room at a very reasonable price.
• Rodeway Inn and Suites Convention Center Los Angeles, 1904 W. Olympic Blvd., 213-380-9393, [57]. Welcome to Rodeway Inn Convention Center Los Angeles. Our reasonably priced motel is ideal for a variety of travelers, and you'll find ample entertainment and attractions in and around our downtown neighborhood.
Budget
• The Vagabond Inn USC Hotel, 3101 South Figuera St, +1-213-746-1531, [58]. Close to the campus of USC. Prices start at $85/night.
Mid-range
• Clarion, 1901 West Olympic Blvd, +1-213-385-7141, [59]. This hotel is conveniently located one mile from the Los Angeles Convention Center and the Staples Center.
• Kyoto Grand Hotel & Gardens, 120 South Los Angeles St, +1-213-629-1200, [60]. A Japanese style hotel in Little Tokyo. Rooms starting at $110.
• Marriott, 333 South Figueroa St, +1-213-617-1133, [61]. Across the street, has a small older theater in its basement where you can still see first-run movies for under $8.
• Omni, 251 South Olive St (at California Plaza in Bunker Hill), +1-213-617-3300, [62]. The OJ jury was sequestered here. Convenient to MOCA, Disney Concert Hall, upscale Noe Restaurant and Bar is onsite.
• Sheraton Hotel, 711 South Hope St, +1-866-716-8130, [63].
• The Standard, 550 South Flower St, +1-213-892-8080, [64]. A very hip and trendy hotel with designer rooms and a bar and swimming pool on the roof. There is also a Hollywood location. Price starts at US$99/night.
Splurge
• Figueroa Hotel, 939 South Figueroa St, +1-213-627-8971 (toll free: 1-800-421-9092), [65]. For those looking for something unusual, Figueroa Hotel provides Moroccan styled luxury. Mystic and beautiful, this is where Cirque Du Soleil hosted their premiere party of Varekai.
• Hilton Checkers, 535 South Grand Ave (located adjacent to the Central Library and the Millennium Biltmore in the Financial District), +1-213-624-0000 (toll free: +1-800-HILTONS), [66]. AAA Four diamond, renovated 1920s luxury hotel. Weekend packages are offered.
• Millennium Biltmore, 506 South Grand Ave, +1-213-624-1011, [67]. The grand-daddy of all downtown hotels, with its gorgeous lobby and fancy restaurants.
• Miyako Hotel, 328 East 1st St, +1-213-617-2000, [68]. A downtown Japanese hotel offering classic comfort accommodations, meeting rooms, and a health spa with Shiatsu massage, close to attractions.
• Westin Bonaventure, 404 South Figueroa St, +1-213-624-1000, [69]. Recognizable from various movies it has appeared in.
Stay safe
The area bounded by 3rd Street, 7th Street, Alameda Street and Main Street is often referred to as "Skid Row" or "the Nickel" and has one of the largest homeless populations in the United States. The Greyhound Station is located here, but the area is unsafe for pedestrians regardless of the time of day.
Contact
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Southern Africa
From Wikitravel
Africa : Southern Africa
Revision as of 06:08, 22 March 2010 by Burmesedays (Talk | contribs)
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Wildlife is abundant in Southern Africa
Southern Africa is a region that includes several non-tropical countries in Africa.
Countries
Botswana (Kalahari, Okavango-Chobe, Southeast)
Famous for the rich wildlife of the Okavango Delta.
Lesotho
The Kingdom in the Sky
Mozambique (Northern Mozambique, Central Mozambique, Southern Mozambique)
Namibia (Caprivi, Northern Namibia, Central Namibia, Southern Namibia)
Namibia produces some of the world's highest quality diamonds.
South Africa (Eastern Cape, Free State, Gauteng, KwaZulu-Natal, Limpopo, Mpumalanga, North West, Northern Cape, Western Cape)
One of the richest countries in Africa has its share of wildlife and tourist-friendly sites.
Swaziland
Zambia (Central Zambia, Copperbelt, Eastern Zambia, Northern Plateau, Zambezi Basin)
Zimbabwe (Matabeleland, Mashonaland, Eastern Highlands, Lake Kariba and the Lower Zambezi, Southeastern Zimbabwe)
Access to Victoria Falls, one of the largest waterfalls in the world.
A number of other countries are sometimes considered part of Southern Africa due to their accessibility from the countries listed above, such as Angola, Malawi and some Islands of the Indian Ocean such as the Seychelles and Mauritius. In some instances all of the countries south of the equator are viewed as Southern Africa. For the purpose of this guide, these countries are described in other regions.
Cities
Other destinations
Understand
Landscape
The Drakensberg (mountains of the dragon) range stretches some 1000km through Southern Africa, from the Eastern Cape through Lesotho (most of this country is on a plateau of the mountain range) , central KwaZulu-Natal (where one can find the Tugela Falls, the second highest waterfall in the world), divides Mpumalanga in half creating the Escarpment and the Blyde River Canyon (the third largest canyon in the world) from where it reaches up into the southern parts of Limpopo.
Talk
English is an official language in all countries in the region, and most urban dwellers speak it fairly well. Afrikaans is widely spoken in South Africa and Namibia, and German is also spoken by some in Namibia. You may encounter Portuguese in the region as well.
Get in
By plane
South Africa's OR Tambo Internatonal Airport in Johannesburg and Cape Town International Airport are two of the easiest entry points into the region, with many direct international flight landing there from Amsterdam, Bangkok,Lisbon, London, Paris, Frankfurt, Munich, Zurich, Athens, Dubai, Doha, New York, Atlanta, Washington, D.C., Buenos Aires, Mumbai, Hong Kong, Kuala Lumpur, Singapore, Sydney, Perth and others.
Connections to the rest of the Southern African region is easily made from here with flights to Blantyre, Cairo, Gaborone, Dar es Salaam, Harare, Lilongwe, Livingstone, Luanda, Lusaka, Kinshasa, Maputo, Manzini, Maun, Mauritius, Nairobi, Victoria Falls, Windhoek and more.
You may also want to have a look at Discount airlines in Africa and Air travel in South Africa.
Get around
See
Itineraries
Trip starts in Zambia, near world-famous Victoria Falls and provides a glimpse into the best of Botswana with plenty of options for game drives before heading down through the wild Limpopo province of South Africa.
Livingstone, Botswana, Waterberg National Park, Johannesburg.
Do
Safari
One of the most popular holiday activities in Southern Africa is wildlife safaris in search of spotting the Big Five (lion, elephant, buffalo, leopard, rhino). There are game reserves within South Africa, Botswana, Zambia, Namibia and Zimbabwe. They all have lodges ranging from basic camping to five-star luxury.
Beaches
South Africa has beautiful beaches stretching all the way from Cape Town around the coast up to Durban, which is a surfing mecca. Further north Mozambique - known for its excellent diving and warm, clear waters - takes over.
Adventure activities
Southern Africa is an adventure haven. The region has some of the highest bungee jumping spots in the world; fantastic hiking and biking trails, great rivers for white-water river rafting and canoeing and excellent surf for surfing and kite-boarding.
Wine
The area at the tip of Southern Africa around Cape Town is known as the Wine Route and produces award-winning wines. Most wine farms are open for public tastings. Some of the towns to visit on the Wine Route include Stellenbosch, Franschoek, Paarl and Robertson.
Eat
Drink
Stay safe
Stay healthy
Water
Depending on the country you are visiting, tap water might not always be safe to drink. Consult the page for the relevant country and check with locals when in doubt.
Sunburn
Many activities in Southern Africa will be outdoors outdoors, see the sunburn and sun protection travel topic for tips on how to protect yourself.
HIV and AIDS
Southern Africa has a very high HIV infection rate.
For your own safety, DO NOT HAVE UNPROTECTED SEX.
Malaria
There might be a risk of contracting Malaria in many areas in the region. See the Malaria and Mosquitoes travel topics for more information.
Get out
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
1304.3 - Monthly Summary of Statistics, Queensland, May 2002
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 30/04/2002
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Australian Bureau of Statistics
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ABS Home > Statistics > By Release Date
1504.0 - Methodological News, Sep 2007
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How I Get A 35% Conversion Rate On My Blog
Posted by GabeJohansson under Online Marketing
From http://gabejohansson.com 87 days ago
Made Hot by: centrifugePR on February 21, 2013 7:44 pm
If you’re struggling to get a good conversion rate on your blog, it’s because not everyone who comes to your site is ready to take action. That’s okay. Inside this video, I reveal a couple tips on how I drastically increased my lead conversion rate on my blog with ease.
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Central African mangroves
Ecoregions:
Central African mangroves
Ndian River Delta, Cameroon. Photograph by WWF/ Mauri Rautkari
This article has been reviewed by the following Topic Editors: Mark McGinley, C Michael Hogan
The Central African Mangroves flank the coastline of western and central Africa, in suitable low energy marine environments. The largest mangrove stand is found in the Niger Delta, which supports the most extensive area of mangrove in Africa. The mangroves of this region have no endemic species but support some endangered species, such as manatees and pygmy hippopotamuses in the Niger Delta. Mangroves are important as nursery and feeding areas for marine fishes, and they trap large amounts of sediment. The oil industry, clearance for salt pans, and overcutting by an increasing human population pose serious threats to these mangroves, but some are contained within protected areas.
Location and General Description
The Central African mangrove ecoregion is located in western Africa, and encompasses mangrove areas along the coastlines of Ghana, Nigeria, Cameroon, Equatorial Guinea, Gabon, Democratic Republic of Congo (DRC), and Angola (to 19°18' S). The structure of the mangrove areas varies considerably, from the lagoon systems found in the western part of this ecoregion to systems modified by complex patterns of sediment deposition at river mouths in the central and southern portions.
Climatic conditions are primarily humid and tropical, but change to more temperate conditions towards Angola. Off the coast of the DRC, mangrove development is inhibited by the presence of the coldwater Benguela Current, but some stands are found where high river water temperatures counteract this current. Annual rainfall varies from a mean of 750 millimeters (mm) in Angola to 6000 mm in Cameroon. In Ghana and the western part of Nigeria, mangroves are primarily associated with extensive lagoons. These are enclosed part of the year by sediments, when rainfall is lower and freshwater outflow is not sufficient to counteract ocean swells. In the remainder of the region, mangroves are primarily associated with river mouths, the largest of which is the Niger River Delta, which may discharge up to 21,800 cubic metres per second at peak flow in mid-October. The sediment load flowing from the Niger River has been estimated to be 20 million cubic metres per annum, most of which is captured in the mangrove swamps. Sediment deposition and channel erosion have created a network of river creeks, estuarine swamps and barrier islands. Soils range from recently deposited unconsolidated, soft dark mud containing silt, clay and peaty clay, to transitional swamps, all of which are associated with different types of vegetation.
The key factors that influence these mangrove ecosystems are river floods and the tidal range. Tidal range increases from west to east, reaching a maximum of 2.8 meters in eastern Nigeria. This allows flood tides to penetrate up to 40–45 kilometers into the interior. The large inputs of freshwater create a low-salinity zone offshore where salinity fluctuations range between zero and 0.5 percent during the rainy season, and 30 to 35 percent during the dry season. Farther south in Cameroon, annual rainfall reaches 6000 millimeters, but is highly changeable because of variation in topography and coastal types. These high freshwater inputs, together with a convergence of the Guinean counter-current, the Benguela current, and an equatorial subsurface current, creates a "piling up" of water that results in an increase in mean sea level of 1.2 m and creates an unusual circulation pattern. It also results in the formation of sandbars and the deposition of large amounts of suspended sediment in the mouths of estuaries.
Five species of mangroves in three families are found in this region, including the Red Mangroves, Rhizophora racemosa, R. mangle, and R. harrisonii, Black Mangrove (Avicennia germinans) and White Mangrove (Laguncularia racemosa), as well as an introduced species, Nypa fruticans. Rhizophora racemosa is the primary colonist in the open lagoon systems, whereas Avicennia africana is the primary colonist in closed systems. Vegetation varies depending on whether the soils consist of sandy troughs or muddy hollows. In the back swamps Nypa fruticans is replacing red mangroves because it is a quicker colonizer and has shallow roots which destabilizes river banks. This is occurring rapidly from the western shores of Nigeria to the interior creeks of the Niger Delta. Rhizophora racemosa is dominant in the tidal and more inundated areas of Cameroon, where mangroves are found concentrated in two locations to either side of Mount Cameroon. Farther south in DRC where mangroves are found around lagoons, the dominant species is Rhizophora mucronata. In Angola, large mangrove communities occur at the mouths of the Cuvo, Longa, Cuanza, Dande, and M'Bridge Rivers, though they are not as extensive as the vast mangrove swamps at the mouth of the Zaire River. The dominant trees are Rhizophora racemosa, R. mangle, R. harrisonii and, Avicennia africana, the former two species reaching heights of approximately 30 meters (m).
Biodiversity Features
While these mangroves contain no endemic species, they are known for their diverse pelagic fish communities, including some narrowly distributed species, abundant avifauna, and the presence of some rare mammals and turtles.
The mangroves provide habitat to the threatened West African Manatee (Trichechus senegalensis, VU), the African Softshell Turtle (Trionyx triunguis), and in the Niger Delta, isolated populations of Pygmy Hippopotamus (Hexaprotodon liberiensis heslopi, VU). The near-endemic Sclater's Monkey (Cercopithecus sclateri) may also use the brackish portion of the mangroves. Hippopotamus (Hippopotamus amphibius), Nile crocodiles (Crocodylus niloticus) and a variety of amphibians are also found in the rivers, lakes and floodplains surrounding the ecoregion. The mangrove forests are also home to the talapoin monkey (Miopithecus talapoin). Summer visitors include at least five species of marine turtle, Leatherback Sea Turtle (Dermochelys coricea, EN), Loggerhead Sea Turtle (Caretta caretta, EN), Olive Ridley Turtle (Lepidochelys olivacea, EN), Hawksbill Sea Turtle (Eretomychelys imbricata, CR), and Green Turtle (Chelonia mydas, EN).
Coastal mangroves and wetlands are primarily important for large concentrations of birds that use the areas during migration, although some wetland species also breed here. Several of the coastal wetland sites are internationally important for migratory wetland birds. The importance of the coastal lagoons of Ghana for wetland birds is summarized by Piersma and Ntiamoa-Baidu, and for the Niger Delta in Hughes and Hughes. Jones also provides a general summary of the biodiversity values of the Gulf of Guinea.
The mangroves are also important to species found primarily in adjacent habitats, but who may also depend on mangroves for parts of their life cycle. The Niger Delta provides spawning/nursery areas for the fisheries in the Gulf of Guinea. A high diversity is found in the pelagic fish community, with 48 species in 38 families. Pelagic families and species associated with them include Clupeidae (Ethmalosafimbriata, Pellonulaleonensis, Ilishaafricana, Sardinella maderensis), Belonnidae (Ablennes hians, Strongylura senegalensis), Megalopidae (Megalops atlanticus), Hemiramphidae (Hyporhamphuspicarti), Elopidae (Elops lacerta, E. senegalensis), and Albulidae (Albula vulpes).
Current Status
Estimates of mangrove area provided by Spalding et al. range between 16,673 and 17,176 kilometers2, of which more than two-thirds are found in Nigeria. Delineations on maps of mangrove areas suggest even larger extents, but estimates are problematic because the mangroves are interspersed with swamp forests.
The most important remaining blocks of habitat are found in the Niger River Delta in Nigeria, to the east of the mouth of the Cross River in Nigeria and Cameroon, around Doula in Cameroon, and the Muni Estuary and Como River in Gabon. Smaller areas of habitat are also found in Ghana, in the Conkouati lagoons of Congo, at the mouth of the Congo River in the Democratic Republic of Congo, and in Angola. The Niger Delta has been growing for millions of years and is still in the process of expanding into the Gulf of Guinea. The delta mangroves mark the transition between swamp forest habitats to pioneer communities on the coast and can extend up to 40–45 km wide.
It is believed that 3,165 km2, or 10.68%, of the Central African mangroves [116], are in some form of Protected Area. These include the Douala-Edea Faunal Reserve in Cameroon (1600 km2) and the Anlo-Keta Lagoon Complex and Songor Lagoon in Ghana. Draft management recommendations have been prepared for the Ghanaian coastal zone, including mangrove areas.
Types and Severity of Threats
Fragmentation itself does not greatly affect mangrove biodiversity, as mangroves are naturally fragmented, and are able to disperse over long distances. Of greater concern is the total amount of mangrove area lost to urbanization, industrialization, and agriculture, as well as impacts from timber and petroleum exploitation. Timber is primarily used for fuelwood and poles for housing construction. Impacts from petroleum exploitation include coastal subsidence that may aggravate the effects of sea-level rise, as well as infrastructure development and oil spills that have led to large mortalities of invertebrates and fishes. Exporting oil from coastal areas is an economically important activity in Nigeria, Gabon, and Cameroon that can lead to oil spills. In Nigeria, during the past 30 years, seismic lines have been placed in the Niger Delta mangrove forests. Other threats include the practice of gas flaring, the use of poison and dynamite for fishing, canalization, discharge of sewage and other pollutants, siltation, sand mining, erosion, construction of embankments, and growing population pressure in the coastal zone.
Justification of Ecoregion Delineation
The Central African mangroves range from Ghana east of the Dahomey Gap, through the Niger Delta (the largest concentration of mangroves in Africa) south to the mouth of the Congo River, with outlying patches in Angola. This ecoregion generally follows that part of the African coastline that is affected (at least occasionally) by the cold water Benguela current.
Further Reading
• Adegbehin, J.O., 1993. Mangroves in Nigeria. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Agyepong, G.T.K., P.W.K Yankson, and Y. Ntiamoa-Baidu, 1990. Coastal Zone Indicative Management Plan. E.P.C., Accra.
• Ajao, E.A. 1993. Mangrove ecosystems in Nigeria. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Appolinaire, Z. Mangroves of Cameroun. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Commission of the European Communities, Directorate-General for Development. 1992. Mangroves of Africa and Madagascar. Office for Official Publications of the European Communities. Brussels, Luxembourg. ISBN: 9282639843
• Diop, E.S., editor. 1993. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Elijah, A. 2001. Strategies for the sustainable management of mangrove resources in the Niger Delta. Discussion paper.
• Hughes R.H. and J.S. Hughes. 1992. A Directory of African Wetlands. IUCN, Gland Switzerland and Cambridge UK/ UNEP, Nairobi, Kenya/ WCMC, Cambridge, UK. Isebor, C.E. and L.F. Awosika, 1993. Nigerian Mangrove Resources, Status and Management. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3. ISBN: 2880329493
• Jones, P.J. 1994. Biodiversity in the Gulf of Guinea: an Overview. Biodiversity and Conservation 3: 772-784
• Makaya, J.F. 1993. Mangroves in Congo. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Niger Delta Environment Survey 1997. Niger Delta Environmental Survey: final report. Vols I-IV. Environmental Resources Managers, Lagos.
• Piersma, T. and Y. Ntiamoa-Baidu, 1995. Waterbird ecology and the management of coastal wetlands in Ghana. NIOZ Report 1995-96, Texel.
• Sackey, I., Laing, E. and J.K. Adomako. 1993. Status of the Mangroves of Ghana. In E.D. Diop, editor. Conservation and Sustainable Utilization of Mangrove Forests in Latin America and Africa Regions. Part II – Africa. International Society for Mangrove Ecosystems and Coastal marine Project of UNESCO. Mangrove Ecosystems Technical Reports volume 3.
• Shumway, C.A. 1999. Forgotten Waters: freshwater and marine ecosystems in africa. Stretegies for biodiversity conservation and sustainable development. The Biodiversity Support Program, Boston University, New England Aquarium, U.S. Agency for International Development.
• Spalding, M.D., F. Blasco, and C.D. Field, editors. 1997. World Mangrove Atlas. The International Society for Mangrove Ecosystems, Okinawa, Japan.
Disclaimer: This article contains certain information that was originally published by the World Wildlife Fund. Topic editors and authors for the Encyclopedia of Earth have edited its content and added new information. The use of information from the World Wildlife Fund should not be construed as support for or endorsement by that organization for any new information added by EoE personnel, or for any editing of the original content.
Citation
World Wildlife Fund (Lead Author);Mark McGinley, C Michael Hogan (Topic Editor) "Central African mangroves". In: Encyclopedia of Earth. Eds. Cutler J. Cleveland (Washington, D.C.: Environmental Information Coalition, National Council for Science and the Environment). [First published in the Encyclopedia of Earth April 17, 2007; Last revised Date March 29, 2013; Retrieved May 18, 2013 <http://www.eoearth.org/articles/view/150996/Angola/?topic=49597>
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Known worldwide by its panda logo, World Wildlife Fund (WWF) leads international efforts to protect endangered species and their habitats. Now in its fifth decade, WWF works in more than 100 countries around the globe to conserve the diversity of life on Earth. With nearly 1.2 million members in the U.S. and another 4 million worldwide, WWF is the world's largest privately financed conservation organization. WWF directs its conservation efforts toward three global goals: 1) saving endangered ... (Full Bio)
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According to this new WHO report India, Indonesia, Pakistan, Myanmar and Papua New Guinea bear the largest burden of malaria in the Asia-Pacific region & account for 89% of all malaria cases in the region.
More research is needed to understand how ecosystems can help reduce disaster risks around the world, according to a report launched in Brussels, Belgium, today.
This FAO study analyses national legislation on wildlife management in 12 countries in Asia and Oceania. It identifies strengths and weaknesses of legal frameworks in promoting sustainable wildlife management and proposes options to support empowerment of the poor.
This publication reports on the human and economic losses caused by natural disasters in 2008 with figures based on the EM-DAT database with comparisons to previous years. It asserts that although fewer disasters occurred in 2008, events had a larger impact than usual on human settlements. In 2008 once again, large numbers of persons were affected by a few natural disasters.
This report provides an overview of information on the world fertilizer situation in 2007/08 and a forecast till 2011/12. The fertilizer situation is examined in relation to crop production and factors likely to affect the latter. High commodity prices experienced over recent years led to increased production and correspondingly greater fertilizer consumption as reflected in tight markets and higher fertilizer prices at the start of the outlook period. While demand for basic food crops, for high value crops such as fruit and vegetables, for animal products and for crops capable of being used to produce bio-fuels is likely to remain strong, it is expected that increased fertilizer consumption required to support higher levels of production will be adequately catered for by growing supply world wide during the outlook period.
This manual starts out with a clear exposé of the contextual issues surrounding the water and sanitation crisis in many countries of the world, encompassing legal foundations, entitlements, responsib
This manual starts out with a clear expose of the contextual issues surrounding the water and sanitation crisis in many countries of the world, encompassing legal foundations, entitlements, responsibi
The main aim of this study was to facilitate access to comprehensive and comparable information on the current and past extent of mangroves in the 124 countries and areas where mangroves are known to exist,
FRA 2005 examines current status and recent trends for about 40 variables, covering the extent, condition, uses and values of forests and other wooded land, with the aim of assessing all benefits from forest resources. In the main section of this report, results are presented according to six themes representing important elements of sustainable forest management: extent of forest resources, biological diversity, forest health and vitality, productive functions of forest resources, protective functions of forest resources, and socio-economic functions.
According to this latest global report on wind energy, total worldwide installations in 2008 were more than 27,000 MW, dominated by the three main markets in Europe, North America and Asia. The United States passed Germany to become world #1 in wind power installations, and China
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Bibliography: The Two Voices
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Hope Vale Aboriginal Shire Council
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Activity Not Available
Contributors : unclewerner
Analyzed about 2 years ago based on code collected about 2 years ago.
Activity on Konzept by unclewerner (see full profile for unclewerner)
All-time Commits: 1
12-Month Commits:
30-Day Commits:
Overall Kudo Rank:
First Commit: 17-May-2009
Last Commit: 17-May-2009
Names in SCM: Frank Mertens, Frank Mertens
Commit history:
Recent Kudos...
... for Konzept given by:
There are no kudos for this contributor at this time.
Project Commits
Approximately one year of commit activity shown
Project Languages
Language Aggregate Coding Time Total Commits Total Lines Changed Comment Ratio
XML 1m 1 7,415 0.0%
Python 1m 1 4,942 9.2%
Make 1m 1 41 -
shell script 1m 1 5 -
All Languages 1m 1 12,403 3.9%
Copyright © 2013 Black Duck Software, Inc. and its contributors, Some Rights Reserved. Unless otherwise marked, this work is licensed under a Creative Commons Attribution 3.0 Unported License . Ohloh ® and the Ohloh logo are trademarks of Black Duck Software, Inc. in the United States and/or other jurisdictions. All other trademarks are the property of their respective holders.
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Contributors
Analyzed about 21 hours ago based on code collected about 21 hours ago.
Commits by Top Contributors
Number of Contributors
Newest Contributors
Name Commits First Commit
leese.thomas81 10 about 3 years ago
muzerakascooby 11 almost 4 years ago
tomman71 16 almost 4 years ago
[anonymous] 1 almost 4 years ago
Top Contributors
Name Kudos 12 Month Commits All Time Commits 5 Year Trend Primary Language First Commit Last Commit
leese.thomas81 0 10 C++ about 3 years ago about 3 years ago
tomman71 0 16 C++ almost 4 years ago almost 4 years ago
muzerakascooby 0 11 C++ almost 4 years ago almost 4 years ago
[anonymous] 0 1 almost 4 years ago almost 4 years ago
See All Contributors
Copyright © 2013 Black Duck Software, Inc. and its contributors, Some Rights Reserved. Unless otherwise marked, this work is licensed under a Creative Commons Attribution 3.0 Unported License . Ohloh ® and the Ohloh logo are trademarks of Black Duck Software, Inc. in the United States and/or other jurisdictions. All other trademarks are the property of their respective holders.
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User:Elizabeth Polidan
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===Fun Links===
===Fun Links===
Revision as of 01:05, 12 February 2013
Elizabeth Polidan
BIOL 398.03 / MATH 388
• Loyola Marymount University
• Los Angeles, CA, USA
Elizabeth Polidan Home
Course Home
Contents
Education
• 2015, BS, Biomathematics, Loyola Marymount University
Course work
• Biochemistry & Cell Physiology
• Conservation Biology Seminar
• Principles of Evolution
• Probability & Statistics
• Biomathematical Modeling (current)
• Biomathematics Research
• Data Structures
• Unix and shell programming
• C Programming
Research interests
1. Stochastic modeling
2. Agent-based modeling
3. Energy budgets
I have been working with Professor Fitzpatrick on a model of the energy budget of a spider.
My Interests in Biology and Mathematics
• My favorite aspect of biology is the fact that everything is connected and inter-dependent. I love the complexity.
• My favorite aspect of mathematics is that you can use it as a tool to understand the world, from ecosystems to social interactions to physics.
Career Interests
Work somewhere to apply my analytical skills to a variety of problems.
Concerns and Information of Note
1. My one concern is that I had cell biology and biochemistry a loooong time ago. I hope that does not affect my ability to understand the biology content.
2. I do not know of anything else I would like the instructors to know. Except, perhaps, that I'm excited about this course!
My Links
Course Homepage
Course Links
Assignments
Week 1
Week 2
Week 3
Week 4
Week 5
Journals
Individual
Elizabeth Polidan Week 2
Elizabeth Polidan Week 3
Elizabeth Polidan Week 4
Elizabeth Polidan Week 5
Shared
Shared Journal Week 1
Shared Journal Week 2
Shared Journal Week 3
Shared Journal Week 4
Shared Journal Week 5
Fun Links
Earth Science Picture of the Day xkcd
Useful links
Files
Homework 1 Notes
Miscellaneous
Personal tools
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User:JeffreyLau
From OpenWetWare
Revision as of 23:54, 14 June 2006 by JeffreyLau (Talk | contribs)
Jump to: navigation, search
About
Undergraduate concentrating in Computer Science (class of 2007), member of 2006 Harvard iGem team.
2006/06/14
Biobrick tutorial
More detailed procedure notes are in lab notebook.
• Began with 12 samples (3x each of negative control, R0010, E0241, E7104). Fortunately our negative controls showed no growth overnight.
• Made 9 minipreps for the remaining samples. Set aside the E7104 samples (they are a positive control).
• Performed digests on 2x each of R0010 and E0241. We lost one of our E0241 samples by accident, so we threw out an R0010 sample to balance.
• Added Calf Intestinal Phosphatase (CIP) to R0010 samples to prevent self-ligation of the R0010 vector.
• Performed gel electrophoresis on R0010 and E0241 samples.
• Our e-gel was not entirely successful.
Must be more careful next time.
Notes on using the nanodrop
• Nanodrop measures concentration of DNA in small samples (1 µL). It is apparently not very accurate.
• Clean nanodrop thoroughly with kimwipes and ethanol, before and after each use
• On desktop, "ND-1000"->"Nucleic Acid"
• Before each set of samples:
• Calibrate nanodrop with drop of distilled water
• Calibrate nanodrop with drop of background (solute), e.g. distilled water or elution buffer
• Add sample to nanodrop, enter name of sample, "Measure"
• Note: mix sample before placing on nanodrop
Personal tools
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Premier League to use newly-approved goal-line technology
PanARMENIAN.Net - Goal-line technology will be used in the English Premier League "as soon as is practically possible" after it was approved by the body that sets the Laws of the Game, the league has said, according to RIA Novosti.
The controversial technology, which will see referees overruled by machines for the first time, will also be used at the 2014 World Cup in Brazil.
The move follows cases of referees allowing so-called "ghost goals" or disallowing seemingly legitimate strikes, as when England's Frank Lampard saw his goal against Germany ruled out at the 2010 World Cup, or when a similar fate befell Ukraine's Marko Devic against England at Euro 2012.
On Thursday the International Football Association Board unanimously approved the use of two goal-line technology systems and FIFA said the technology would be showcased at the Club World Cup in December.
The Danish-German GoalRef system uses magnetic fields to determine whether the ball has crossed the line, while Hawk-Eye uses cameras and is widely used in tennis, cricket and volleyball. Both systems passed FIFA testing in March.
"The Premier League has been a long-term advocate of goal line technology," the league said in a website statement.
"We will engage in discussions with both Hawk-Eye and GoalRef in the near future with a view to introducing goal-line technology as soon as is practically possible."
FIFA president Sepp Blatter said he would push for its inclusion at the World Cup in Brazil.
"It is my plan to have it used at Brazil 2014, yes. We will use the system also for the Confederations Cup and this year’s FIFA Club World Cup," he said on the FIFA website.
The board also approved continuing to use extra assistant referees on the goal line, a scheme championed by Michel Platini.
The additional assistants, introduced in 2009, came in for criticism at Euro 2012 after one failed to spot that Devic's strike had crossed the line.
Partner news
Top stories
Jorge Rafael Videla, an austere former army commander, led Argentina during the bloodiest days of its Dirty War dictatorship.
According to the United Nations, April was Iraq's bloodiest month for almost five years, with 712 people killed.
Reports suggest the rebel fighters may have tried to blow up the walls of the prison, which holds some 4,000 inmates.
Moscow has condemned other nations for supporting rebel forces and failing to condemn what it describes as terrorist attacks on the Syrian regime.
Partner news
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Sunday, June 19, 2011
Last statement sent to Sentinel from self-immolation victim
SentinelSource.com
S.Paul Note: This is reminiscent of the man who flew his plane into an IRS building not too long ago. Similarly, he felt there was no way out of the strangle hold on him by a system gone wrong. Those who support this system have labeled both men insane or deeply troubled in order to deny the intrinsic sickness created by an insane system. When I read the last line, I began to become misty and had to fight off grief for not only he but his children and all those others in this world who have fallen victim to the corruption of an authority lost within a broken system that no longer views We the People as a worthwhile investment.
Sentinel Editor's Note: On Thursday morning, June 16, The Sentinel received a "last statement" via mail from a man who insinuated that he planned to set himself on fire in front of the Cheshire County Court House, and an explanation of why he intended to do so. Through further reporting, The Sentinel is confident this is from the victim of Wednesday afternoon's fire, although police have not yet received confirmation of his identity. The 15-page statement is printed in full, except for two redacted items: The names of the man's mother and his three children.
Last Statement
by Tom Ball: May he rest in Peace
A man walks up to the main door of the Keene N.H. County Courthouse, douses himself with gasoline and lights a match. And everyone wants to know why.
Apparently the old general was right. Death is not the worst of evil.
I am due in court the end of the month. The ex-wife lawyer wants me jailed for back child support. The amount ranges from $2,200. to $3,000. depending on who you ask. Not big money after being separated over ten years and unemployed for the last two. But I do owe it. If I show up for court without the money and the lawyer say jail, then the judge will have the bailiff take me into custody. There really are no surprises on how the system works once you know how it actually works. And it does not work anything like they taught you in high school history or civics class.
I could have made a phone call or two and borrowed the money. But I am done being bullied for being a man. I cannot believe these people in Washington are so stupid to think they can govern Americans with an iron fist. Twenty-five years ago, the federal government declared war on men. It is time now to see how committed they are to their cause. It is time, boys, to give them a taste of war.
There are two kinds of bureaucrats you need to know; the ones that say and the ones that do. The bridge between them is something I call The Second Set of Books. I have some figures of the success of their labors. You and I are in these numbers, as well as our spouses and children. But first let me tell you how I ended up in this rabbit hole.
My story starts with the infamous slapping incident of April 2001. While putting my four year old daughter to bed, she began licking my hand. After giving her three verbal warnings I slapped her. She got a cut lip. My wife asked me to leave to calm things down.
When I returned hours later, my wife said the police were by and said I could not stay there that night. The next day the police came by my work and arrested me, booked me, and then returned me to work. Later on Peter, the parts manager, asked me if I and the old lady would be able to work this out. I told him no. I could not figure out why she had called the police. And bail condition prevented me from asking her. So I no longer trusted her judgment.
After six months of me not lifting a finger to save this marriage, she filed for divorce. Almost two years after the incident, I was talking with her on the phone. She told me that night she had called a mental health provider we had for one of the kids. Wendy, the counselor told my then wife that if she did not call the police on me, then she too would be arrested.
Suddenly, everything made sense. She is the type that believes that people in authority actually know what they are talking about. If both she and I were arrested, what would happen to our three children, ages 7,4 and 1? They would end up in State custody. So my wife called the police on her husband to protect the children. And who was she protecting the kids from? Not her husband, the father of these children. She was protecting them from the State of New Hampshire.
This country is run by idiots.
The police sergeant Freyer screwed this up from the get go. When I got the Court Complaint form the box was checked that said Domestic Violence Related. I could not believe that slapping your child was domestic violence. So I looked up the law. Minor custodial children are exempted. Apparently, 93% of American parents still spank, slap or pinch their children. To this day I still wonder if Freyer would have made this arrest if it had been the mother that had slapped the child.
Labeling someone's action as domestic violence in American in the 21st century is akin to labeling someone a Jew in Germany in the 1930's. The entire legal weight of the state is coming down on him. But I consider myself lucky. My family was destroyed. But that poor bastard in Germany had his family literally annihilated.
Arrests are mandatory for the police in New Hampshire for domestic violence. That is not law. That is police department policy. Laws come from the Legislature and the Governor's office together. God only knows where these policies come from. The State's Attorney General also has a mandatory arrest protocol for domestic violence. I call these policies, procedures and protocols The Second Set of Books. You never cover the Second Set of Books your junior year in high school. That because we are not suppose to have a Second Set of Books. This is America-we have the rule of law.
I am a regular guy, a coffee and cheeseburger type of fellow. As remarkable as my life has been, I figure that what happens to me must be happening to others as well. I was 48 years old when I got arrested here for my first time. So I went looking for the arrest numbers for domestic violence, this new group that I had unwilling joined. I could not find anything. So I wrote the U.S. Dept. of Justice in Washington. They wrote back that they did not keep track of domestic violence arrests. The FBI keeps track of all other crimes. How come not domestic violence? I thought some low level clerk was blowing me off.
At the time, I had mailing addresses in both New Hampshire and Massachusetts. So I wrote to all six Congressional offices, the two Senators from each state and the two Congressman. They like doing favors for constituents hoping you will favorably remember their name in the voting booth. All six offices reported back the same thing. They do not know how many arrests for domestic violence have been made. I immediately knew something was wrong. And I also knew this was not going to be good.
Improvise, adapt and overcome. The Army teaches that to every soldier it trains. They say that no battle plan survives the first five minutes of combat. So your people on the ground had better be able to think for themselves. Taking casualties in war is just an occupational hazard. Taking casualties and not accomplishing your mission is a disaster. After 21 years of Army service, I am pretty good at improvising.
The first thing I found was a study not of domestic violence arrests but of domestic violence injuries for 18 unnamed states and the D.C. in the year 2000. In the study 51% of the injuries were 'no injuries'. So I knew I had a study of police reports. Who else but a police officer would record no injuries? I populated that out to the 50 states and came up with 874,000 arrest in the year 2000.
I had originally populated the number back to 1994 when the Violence Against Women Act (VAWA) was enacted into law. I would later find out these arrests stated with the U.S. Attorney General's Task Force of Domestic Violence ten years earlier in 1984. As individual states data became available for various years and states, I would incorporated in to my informal study. The number I have now in 2011 is 36 million adults have been arrested for domestic violence. I have a gut feeling this number could be as high as 55 million. But I only have data to 36 million. So 36 million it stays. And there is a really cool trick you can do once you have this number. You can find out how many American men. women and children ended up homeless because of these arrests.
Most of the domestic violence statistics I have seen break down with 75% male and 25% female being arrested. So I am going to used the male pronoun for the one arrested spouse and the female pronoun for the victim spouse. That should make the domestic violence feminists ecstatic-man bad, woman good. But that is okay because that is probably the last nice thing I am going to do for them today.
When then a man is arrested for domestic violence, one of two things can happen. If they are only dating and have separate apartments, then he can head home. But if they are living together, then this fellow has a real problem. Bail conditions and then a possible protective or restraining order prevent him from being with her. So he needs to find a new place to live, at least until the charges are resolved. The King of his Castle is no longer allowed into his castle. A feminist name Pence who wrote that was absolutely giddy at that outcome. So he can get his own place if he has enough money. Or he can move in with his mother, his sister or another relative. He might have a girl friend who would let him stay with her. And if none of this is possible, well then I guess he is sleeping in his car down by the river.
If he has minor children, money will soon turn into an issue. Most men I know do not mind paying child support. They want their kids to have food on their plates, clothes on their backs, and a roof over their heads. But it does stress that man's finances. Child support is usually 33% of the man's gross income. Withholding for taxes, social security and health insurance can range up to 28% of his gross paycheck. So a man making $500 a week gross has only $825 monthly left over after withholding and child support. That is not enough money for an apartment here in Central Massachusetts. That does not include other expenses like heating, electric, gas, groceries, telephone, cable, car payment and car insurance. So he is in a financial hole. Estimates of homeless men run 82% to 94%. I am going to round that down to 80%.
After the King has left his castle, his wife runs into a problem. She was use to getting his whole paycheck for the household. Now she get a third for child support. Figure they both work and made the same money, her budget went from 100% down to 66%. If she was running the house on $3,045. a month when the King was home, now without him she only has $2,220. Most households in America cannot withstand a 27% hit on the household account. She'll juggle the bills but eventually most wives figure out that they can pay all the smaller bills if they just does not pay the big bill. That would be the rent or the mortgage. So six to nine months after the King is out of the castle, the Queen, the Princes and the Princesses are also on the street. Domestic violence feminists state that 50% of victim spouses of domestic violence end up homeless at some time in their lives.
The last group of homeless from these arrests are children. The domestic violence feminists state that 70% of domestic violence couple have children. So 50% female times 70% children equals 35%. But children is plural. So we will double to 70%.
(Odd isn't it? They know that 50% of victim spouses end up homeless and that 70% of them have children. How can they know the percentages when they do not know how many total arrests were made? Those people at the U.S. Justice Dept. cannot even pull off a credible cover-up. )
Men are 0.8, women are 0.5, and children are 0.7 for a grand total of 2.0 homeless Americans for every domestic violence arrest. Multiply that by 36 million and you get 72 million men, women and children ending up homeless at some point in their lives over the last 25 years because of these domestic violence arrests.
That is a really large number even by Washington standards. That is almost 25% of the entire population of the U.S. using 2010 census figures. Which begs the question did these homeless people contribute to this latest economic meltdown, or did they cause it? Because if they did cause it then the recovery will not be measure in months or years but in decades.
Some of the boys in the Father's Movement think Congress might have shot themselves in the foot over this one. Personally, I think they shot themselves some place anatomically higher. No wonder the Speaker of the House is always crying. The Dummies on the Potomac.
Twenty-five years ago the federal government start pushing these arrests on state's legal systems. Now, we have an economy on the rope. They have thrown a huge amount of money at banks, big business and local and states government. And we are still in the mud. But no economist either at the Treasury Dept., Federal Reserve, universities or think tanks are even looking at the impact of all these broken families. If that 36 million arrest is correct, then 72 million men and women, have been throw out of the middle class into subsistence living. Or is the number 55 million and 110 million? No one knows and no one is even looking. But why should look? According to the Attorney General, we do not know how many arrests we have made.
And if the Tea Party is any indication, insurrection is brewing in the land. Just a coincident? Not likely. This is what happens when the government wipes out the middle class.
The idea for these arrests came from something called the Minneapolis Police Experiment (MPE) of 1981-82. In the experiment police offices were given pads with one of three words written on them; counsel, send or arrest. Counsel meant the officer was to try to mediate the couple's spat. Send was to send one of the spouses out of the house for eight hours as a cooling off period. Arrest was arrest one of the two spouses. The officer was to do as the top paper on the pad said to do. The experiment was set up by the Police Foundation and Lawrence W. Sherman was the lead researcher. The results show counseling resulted in a future assault in 24% cases, send was 19%, and the arrest option resulted in a future assault in only 10% of the cases. Perhaps a cheap way of cutting down future domestic violence.
In 1984 The U. S. Attorney General's Task Force of Domestic Violence recommended arrest as the primary weapon in domestic violence assault. Lawrence W. Sherman recommend not using the arrests because the MPE was just one study and it could be wrong. They ignored him. And by 1992, 93% of the police departments in the nation had adopted some form of mandatory arrest in domestic violence cases.
But by 1992 five more addition studies similar to the MPE became available. Lawrence W. Sherman reviewed all five studies. Then once again he wrote that the police should not use arrest. In two of the five studies, they found the same result as they did in the MPE, that an arrest cut down the odds of a future assault. But in the other three studies an arrest actually increase the odds of a future assault. So arresting someone in a domestic violence situation to cut down on future assaults did not work any better than just flipping a coin. I do not know if Lawrence W. Sherman is still alive. But fortunately he wrote a book call Policing Domestic Violence that was published in 1992.
So we have 800,000 American police officers arresting one in every six adults in the country and throwing 25% of the men, women and children out on the streets in an effort to enforce a policy that they knew did not work back in1992. And I had always assumed that you needed a man to really screw something up. Oh well, there goes another glass ceiling.
Why would they push an arrest policy that does not work? There are two schools of thought on the reason why. The first comes from Lawrence W. Sherman. He calls it the Law of Just Desserts. Revenge for slights and offenses, real or imagined. I am sure there are some that would argue that women are not vengeful. But what is that old saying? Hell hath no fury.....
The second idea comes from the mother of the second wave of feminism. I do speak of the brilliant Betty Friedan. In the Epilogue Chapter of the 20th Anniversary Edition of her book The Feminine Mystique, Betty relayed why she resigned as the first president of the National Organization of Women in 1970. Betty wrote that she, "was unable to openly fight the man haters and unwilling to front for them any more..." So man hating bigots no only existed 40 years ago, they were also grabbing power. Now Washington is funding them. Makes you wonder what bigots they will fund next. Maybe the Klan?
Feminists had always claimed that when women took over, we would have a kinder, gentler, more nurturing world. After 36 million arrests and 72 million evictions what we got was Joe Stalin.
The third wave of feminists do not like to call themselves feminists. The word feminist could be perceived as gender oppression. These third wave of whatever-we-call-you got that right The treachery of our legal system over the last 25 years may end up giving all feminists a bad name. Which would make us as bigoted as the man-hating feminists who got us into this mess to begin with.
So let us talk about those bureaucrats that do. These are the ones that actually carry out the evil deeds. I like call them the do-bies.
Any one swept up into legal mess is usually astonished at what they see. They cannot believe what the police, prosecutors and judges are doing. It is so blatantly wrong. Well, I can assure you that everything they do is logical and by the book. The confusion you have with them is you both are using different sets of books. You are using the old First Set of Books- the Constitution, the general laws or statutes and the court ruling sometime call Common Law. They are using the newer Second Set of Books. That is the collection of the policy, procedures and protocols. Once you know what set of books everyone is using, then everything they do looks logical and upright. And do not bother trying to argue with me that there is no Second Set of Books. I have my own copies at home. Or at least a good hunk of the important part of it.
I got my Second Set of Books when I sued the Jaffrey NH police department. Under the discovery rule, I write them with the material I wanted and it would arrive in the mail a few weeks later. I got the Police Academy Training Manual. I got the Department's Policy and Procedure Manual. I got the no-drop protocol that the attorney general sent to all his or her prosecutors. I even got the domestic violence protocols for the court system, one hundred pages worth. Once you read it the material, then you will know what the police, prosecutors and judges will do. They are completely predictable once you know what set of books they are using.
The police academy training manual states that an arrest in a domestic violence call is the preferred response. They cite the Minneapolis Police Experiment (MPE) as its justification. But the author of the MPE, Lawrence Sherman, said do not use arrest because five follow up studies show that it did not work. The would be a violation of the 4th Amendment in the First Set of Books against unreasonable search and seizure. Then there is that whole issue of whether the police have the right to arrest for any reason other than they believe a crime was committed.
The Jaffrey Police Department Policy and Procedures Manual states that if a wife says she does not want her husband arrested, the police are to ignore her, arrest the husband, and get with the prosecutor to see what they can work out. In other words, make the arrest and then see if you can Mickey Mouse it. The wife is eligible for spousal immunity. If she invokes it, then no statement she mades, written or oral, are admissible because she cannot be cross examined about it under oath. ( Did you say that? What did you mean when you said that?) With no statements the police have no probable cause in most cases to make an arrest. Also a violation of the 4th amendment in the First Set of Books.
The actor Nickolas Cage was drunk in New Orleans with his wife. Everyone else is drunk in New Orleans, so why should Nick be any different. He and his wife were arguing over which house the rented for their stay. Nick grab his wife's arm and started to lead her to his house. The police arrested Nick for domestic violence. His wife was stunned. That was not domestic violence. "Nothing we can do," the police explained to her. "Just following orders."
That is an accurate explanation for victims, even if they do not think of themselves as victims. The police have a zero tolerance towards any physical contact. Things might get worse in the future is the feminist logical for this present iron fist approach to domestic relations. I would have to agree with them. After all the arrests, poverty, homelessness and misery, I can assure you-things are going to get worse.
But that nothing we can do, just following orders the officers explain always sounds so timid and lame. The police need to punch their explanation up a bit, make it more authoritarian. And there is a quick, low cost way of doing it. The police officers only need to say it in its original German.
The state Constitution in NH said the prosecutors job is to promote justice. The Attorney Generals protocols said that domestic violence case are no-drop cases. (Unless, of course, they take the Deal. Continue the case for a year, go to counseling, and everything falls off the books after the year. They did after all find some way of getting rid of all these cases.)
The Attorney General can hire, fire, layoff, promote, demote, commend or award bonuses. The constitution is some old, quaint, dusty document up in the Statehouse some where. So which one do you think is going to get obeyed?
Prosecutors are funny. Some, maybe most, have egos the size of Cape Cod. But of the three, police, prosecutor and judges, prosecutors have the least protection. Micheal Nifong, the prosecutor in the Duke Lacrosse Rape Case, was fired, disbarred, convicted of a crime, and actually jailed for trying to enforce the no-drop prosecution protocol for sexual assault in the Duke case.
The prosecutor in my criminal case fared a little better. I filed a complaint with his boss for summoning my two daughters, ages 7 and 4, to court. I had already conceded that the facts were not in dispute. The trial would be about the law. No witness were need much less a couple of toddlers. He still summoned them. (The Second Set of Books tells the prosecutors to get a sympathetic face in front of the judge or jury. What's more sympathetic than toddlers.) The prosecutor could not refute my allegation because I enclosed a copy of the trial transcript. I had to pay for the transcript. When the prosecutor read it, he gave his two weeks notice and then blew town. That transcript was the best $46 I had ever spent in this life.
There is a name for what happens when a bureaucrat is destroyed by the First Set of Books for attempting to enforce the Second Set of Books. It is called the Abu Ghraib Syndrome. The people within the law enforcement community no longer seem to know the difference between the law, with its checks and balances, and the policies, procedure and protocols that constitute The Second Set of Books. In some cases you do not even know who wrote the policy, procedure or protocol. It could have been the local high school gym teacher for all anyone knows. Many of these bureaucrats are eventually going to learn the different between the First and the Second Set of Books. And my guess is that many of them are going to learn it the hard way. Because the only checks and balances in The Second Set of Books is The First Set of Books.
Judges routinely use our children as bargaining chips. Get the adult into counseling, continue the case for a year, and then drop it. This will open up the docket for the new arrests coming in next week. These judges that use our children are not honorable. Which is why I never use the term 'Your Honor' any more. I just call them judge.
Alex Baldwin, the actor, wrote that you have never seen a coward until you have seen a Los Angeles County judge. I call my judges-Sullivan, Arnold and even Runyon-cowards, too. When I first started observing them, nothing made sense. Arnold was completely infuriated when he was maneuvered into ruling not guilty. He verbally went up and down me so many times I lost count. What was the big deal? If I was not guilty just say and then we could all go home. But that was back in the days before I knew about The Second Set of Books.
I lost visitation with my two daughters when I got arrested. One was the victim-the other was the witness. After a not guilty, I expected to get visitation with my girls. But the divorce judge, Sullivan, decreed that counseling was in order and they would decide when we would reunite. I told the judge that the decision on whether these two girls had a father or a fatherless childhood was not leaving this courthouse. There would be a couple of reason for that decision.
First, by then I knew of the Second Set of Books. As much as I had prayed for the return of my children, I knew that this counseling might get thrown in the way. Judges are addicted to counseling like a meth-head is addict to crystal meth. Sullivan wrote in the divorce decree that he envision only one or two meetings with the counselor. There is no counseling done in the first meeting or two. It is intake-who's the players and what are the issues. But Sullivan was not interested in counseling. He merely wanted to unload the decision out of habit. And if we do not shut them down now, they will be doing it to our kids in twenty years from now when they have little ones running around the house.
Second, just exactly where does the buck stop with our legal system? Police have to make an arrest. The prosecutor has to pursue the case. Judges now also walk a away without rendering a verdict, and passing the buck does not constitute a decision. Can those mental health counselors slide the decision over to someone else? Just where does this end? Who is responsible? Who is accountable?
The mental health crowd is the third reason I said no. Some people think they are geniuses with their Masters and PH D's. Others think they are so wacky that they call them fruit loops. Well, I have a third name for them. Suckers. They did not get hired for their medical ability. They got these because they were willing to take these cases off the judge's hands. Which has done nothing for the credibility for their profession. We are not here to help-we are here to unload. And they created a liability that did not previously existed. If a judge releases a defendant and he goes kills someone, that judge or the judiciary cannot be sued. But a mental health worker, and their employer certainly can be held liable. Our judiciary is now using the mental health field like a ten dollar whore.
I sued Monadnock Family Services to make them go away. I told their lawyer Byron that they were a legitimate target for men. We settled for no money. They would have nothing to do with this reconciliation. The counselor was released. And they would no longer get involved in any domestic violence cases.
Every time we ended back in court over whatever squabbles, I would ask Judge Sullivan for my children back. The decision belong to the counselors he would tell me. But he knew he had screwed up. I could see it in his face. But he would not fix it. He would not step out of that box those domestic violence/sexual assault advocates had built for him. After five years, he retired to a part time position at the Littleton courthouse 120 miles away.
So when guys like Alex Baldwin and I call judges cowards, we have legitimate reasons for doing so. It is not good for judges to be called coward. It is unlikely that it is good for the rest of us.
I do not claim to have all The Second Set of Book. I know of one book that I do not have. And I would have love to read that one. That would be the seminar that the domestic violence and sexual assault advocates put on periodically for legal personnel including judges. These advocates are camped outside every state, not federal, courthouse in America. The U.S. Dept of Justice provides 50-100% of their funding depending on the program. They have three day seminars at resorts where everything is paid for except the liquor. Judges in NH are ordered to attend. Neither Sullivan or Arnold would confirm or deny they had attended. They actually said nothing. It must be like the Masons where they will not say anything about the organization until you show them the secret hand sign.
Supreme Court Judge Louis Brandeis once wrote that the best description of a judge is the impartial guardian of the rule of law. How does three days of wine, women and song contribute to impartiality? It does not. So it should not have been any surprise that they would not answer me. After all, they were not on trial. I was. But they are going to be. They were suppose to protect to rule of law not collaborate in its demise. They have failed miserably.
A guardian ad litem is an attorney appointed for a child. The attorney solely represents the child. I got one when I was first separated to get a neutral pair of eyes and ears on the family. I was disappointed in his findings.
A few years later, another guardian was appointed for one of the kids. A regular report filed with the Court painted me as some sort violent psychopath. I thought that was uncalled for seeing as we had never met. It start a flurry of nasty letters between until we both came to the conclusion that this was not about us. We ended on a friendly note.
At a Court hearing later on I approached him. I asked him if he had had any domestic violence training. He said yes, that it was required to become certified as a guardian ad litem. Another chapter for The Second Set of Books that I never managed to acquire.. So men, if you were thinking about getting a guardian ad litem for an unbiased assessment, then you should ask for the domestic violence material that certified the guardian. And do not worry that you are not sure what you are looking for. It will stand right out.
There are more sections of The Second Set of Books. Medical personnel are supposed to report suspected domestic violence. The college professor Angela Davis has a story of a Latino couple in California getting in trouble feeding the dog his liver for dinner. Mental health employees are also required. Think of Wendy threatening our kids with foster care. Teachers, day care providers, the list just goes on and on. The East German secret police, the Stazi, had 25% of the population on record as informers. The United States is not that high yet, but we are still growing.
These people-police, prosecutors and judges-are suppose to protect us. They are checks and balances to prevent injustice. That is why we spend so much money of police training. But if the police screw it up, the prosecutor can catch it. If the prosecutor misses it then the judge can step in to fix it. But if all three have been compromised, then what does one need to do to get justice? Go to the appeals court or the Supreme Court? That seem a little ridiculous particularly when the zero tolerance has arrests for something as trivial as touching.
On one hand we have the law. On the other hand we have what we are really going to do-the policies, procedures and protocols. The rule of law is dead. Now we have 50 states with legal systems as good as any third world banana republic. Men are demonized and the women and children end up as suffering as well.
So boys, we need to start burning down police stations and courthouses. The Second Set of Books originated in Washington. But the dirty deeds are being carried out by our local police, prosecutors and judges. These are the people we pay good money to protect us and our families. And what do we get for our tax money? Collaborators who are no different than the Vichy of France or the Quislings of Norway during the Second World War. All because they go along to get along. They are an embarrassment, the whole lot of them. And they need to be held accountable. So burn them out.
In the last 25 years they have arrested one in six adults in this country and forced 25% of the men, women and children into homelessness. In 50 years it will be one in three adults arrested and 50% of the men, women and children ending up homeless. Most of our kids will live to the age of 68 years old. As bad as it was for you, your children will have twice the odds of it happening to them.
Some of you will say that 50% homelessness sounds absurd. But 25% is absurd and that is already here. There is no evidence that the police, courts, or government is planning to do anything different in the immediate future. And they will not do anything different until we make it so uncomfortable that they must change. Bureaucracy at its worst. So burn them out. This is too important to be using that touchy-feeling coaching that is so popular with business these days. You need to flatten them, like Wile E. Coyote. They need to be taught never to replace the rule of law. BURN-THEM-OUT!
Most of the police stations built in New England over the last 20 years are stone or brick. Fortunately, the roofs are still wood. The advantage of fire on the roof is that it is above the sprinklers. But even the sprinklers going off work to our advantage. There is no way they can work in a building with six inches of water. And I am certain we will disrupt their momentum once they start working out of a FEMA trailers. If they still do not get the message, then burn down the trailers.
The easiest way of burning a building is with the Molotov cocktail. It was invented by the Finns when the Soviets invaded in 1939. You fill a bottle with gasoline and stuff a rag in the end for a wick. You light the wick and throw bottle, It shatters on impact spraying gas everywhere and the wick ignites the gas. Simple, readily available, and effective. And only two things to remember.
First, use a glass bottle. Thinner glass is better than thicker glass. You want it to shatter on impact. When I was teaching a kid at the high school on the West Side Worcester, MA. threw a Molotov cocktail into his school. Fortunately, he used a plastic bottle. It burned about three square inches of carpeting. I had to laugh when I said to myself, "Thank God for dumb kids."
Second, you need to tie the rag to the bottle. Nothing worse that throwing a Molotov cocktail, landing where you wanted it, and having it shatter perfectly. Then you noticed the wick had fallen out on the way to the target. No wick-no fire.
Some of these building will have brick faces and metal roofs. Just break a window and throw the Molotov cocktail inside. Carpets, furniture, computer plastic, even paint on the walls will burn. It is okay if the sprinkler goes off. I wonder if you can get hip waders over a gun belt?
We had a kid in my hometown that burned down the old junior high school. He walked up to the front door one night with a can of lighter fluid. The applicator on the end squirts the lighter fluid out. He squirted under the door and along the seams and lit a match. The kid took out the entire old part of the building. Why are kids so competent when it is something they should not be doing?
There will be some casualties in this war. Some killed, some wounded, some captured. Some of them will be theirs. Some of the casualties will be ours.
Now, nobody wants to get killed. But let us look at your life. You are broke after paying child support. She and the kids are not doing any better. None of you are middle class any more. You have no say in the kids education, their health treatment, you may not even have visitation with your sons and daughters. And everything you thought you knew to be true-the rule of law, the sanctity of the of the family, the belief that government was there to nurture your brood-all turned out to be a lie. Face it boys, we are no longer fathers. We are just piggy banks.
So you are not losing anything by picking up the Molotov cocktail. It may be too late for us. But without something changing, your kids will have double the odds of it happening to them. That will knock them out of the middle class again, providing they ever get back in. And their kids, your grandchildren, will end up damaged goods before it is over. So it is okay to run. You just need to turn around and run at them. They are no way as imposing as they seem. They only do what they do for a paycheck.
Television would make us believe that people get arrested because of fingerprints, DNA, facial recognition, and instruments that can tell where a substance was made and here is the local distributors. It is Hollywood crap. Most of the people in prison are there for one key reason. They could not keep their mouths shut. They told someone. That someone told others. The cops hear it and start looking at them for a suspect. That how it works in real life.
This need to confess seems to be primeval. Just human nature. But if you cannot keep a secret, do not expect the one you tell to keep their mouth shut. There is only three people I know for certain they will keep their mouths shut. That would be Jesus, Mary and Joseph.
I only managed to get the main door of the Cheshire County Courthouse in Keene, NH. I would appreciate it if some of you boys would finish the job for me. They harmed my children. The place is evil. So take it out
Some where along the line I picked up the crazy notion that it is better to be dead as a free man than to live as a serf. The government needs to be a little more careful about what they teach in our schools.
And bring a can of spray paint to these fires. Paint the word COLLABORATORS ( two L's with an S on the end) on the building before you burn it. Maybe we can shame them back to the rule of law. And we do want the police to know exactly who burned the building. Then the police can start interviewing the usually suspects, all 36 million of us.
We have covered the do-bies. Now let us look at the bureaucrats that say-ers.
The Second Set of Books originated in Office of Violence Against Women (OVW) which is part of the United States Department of Justice. Some of these policies, procedures and protocol were developed locally. But the local results would be sent up to OVW and, if approved, would disperse it out to all 50 states. They are smart, clever, bigoted and able to lie as well as any politician that ever called Washington home. In other words, they have now become Washington insiders.
But what makes them so uniques is their anger towards men, any man. They are so twisted in their hatred of men that they are positively scary. And it is not what they are doing to men that makes them frightening. You would expect that. No, it is what they are doing to the women and children that makes them so twisted.
When the Pentagon drops a bomb on innocent civilians the military calls it Collateral Damage. It sounds better than, "Yeah, we killed a bunch of women and children." Those poor, innocent, stupid civilians have always been caught in the middle since the time we were fighting with rocks.. Your wife and kids are Collateral Damage in the war against you, the man in the family. For 25 years these feminists at OVAW have been willing to sacrifice the women and children to get you. And they cannot claim ignorance about what they are doing. Under the VAWA the federal government is funding at least 1,800 homeless shelters. As long as the Office for Violence Against Women exists in the U.S. Department Justice , no American man, women or child will be safe in their own home.
If you ask these feminists why are the shelters all full, they will not say because of all the arrests. The shelters are full because of men. But they knew from the beginning that this was not man bad-woman good thing. The year was 1976. Two things would happen that year.
First, someone at the U.S. Dept of Justice decided to count the dead bodies. In 1975 there were 1522 women killed in domestic violence. And for men killed in 1975? The dead for men was 1506. Statistically equal a friend tells me so.
If you had asked me before the study, I would have assumed that women were getting the worst of it. But I would be looking at it by genders. What I should have been looking at was species, homo-sapiens, human beings. Men are human-women are human. Being the same species you would expect the same results from both genders. And that is exactly what the dead bodies told us.
The second thing that happened in 1976 was the first domestic violence survey was released. It was so new the time that they called it family violence. Murray Straus of UNH and Richard Gellars from a school in RI were the researchers. They did not find two perpetrators of domestic violence, but three. Men initiated violence 25.7% of the time: women 25.2%, and the other 49.1% was the two going after each at the same time. These two people going after each other at the same time is well recognized in law. The law in NH calls that mutual combat. Men are human. Women are human. And once again we found both genders acting the same manner.
So how did we end up with the theory of man bad-woman good that the government at all levels is using? The feminist writer Susan Brownmiller wrote In Our Time that," the way you get funding and church donations is to talk about the pure victims. If you talk about the impurity of the victim, the sympathy vanishes." If women get to be good then men get what is left-bad. Man bad-woman good was originally a funding raising technique. After 35 years, it has turned into official government dogma at all levels, from the local cop on the beat to the White House. Men need to be punished, restrained and retrained. Your wives and children are, unfortunately, just collateral damage in this effort to punish men. So you were not dreaming it. There really is a government pogrom against men.
When a man batters or kills, there is no excuse. When a woman commits the same act, there is nothing but excuses. Simple though inaccurate. But there is one redeeming aspect to men being demonized. Now we men can act like devils. And we do not even need to apologize for it. Men are going to start acting just like they made us out to be. As an old high school semi-punk I can assure you boys of one thing. This is going to be fun. You guys are going to end up laughing like hyenas.
The money funded under the VAWA is split in two when it leaves the Treasury. Part goes the Health and Human Services for fund these domestic violence homeless shelters. If that 36 million number is correct, and it is all that we have, then the 1.44 million arrests a year will be made producing 2.88 million homeless Americans each year. Women and children constitute 60% of these homeless people, 1.7 million Americans a year. Shutting down these shelters would be cruel. What would these women and children do then? Go live under a bridge. No, we are stuck with these shelters for a while. But there is one thing that Congress needs to fix when they fund them again.
These shelters do not allow men on the property let alone inside the residences. Why is it against the law to use federal money on organizations that discriminate against black, Jews, gays or even women but it is okay to do so against men? Men contributed half that tax money. Eight years ago a man in California fled with his children after the police warned him to get out after they had arrested the wife and mother. None of the shelters would take him and the kids in because he was a man. I wonder if this would survive a legal gender discrimination challenge in a federal court?
A society without men is freakier than a world without blacks or Jews. That is not to say blacks or Jews are any less worthy. It just that there are more men in the world than blacks or Jews even if you combined them. If these feminist had to deal with men on a regular basis, then maybe the country would not be in the pickle we are in now.
There is a third reason to end this discrimination, something of a more practical nature. Apparently, some women like to have sex with men. But men are barred from the property. Suddenly, that 15 year boy two doors down starts looking real good. It might even be fun breaking in this new meat. So this woman driven into insolvency by the push for domestic violence arrests now finds herself charged as a pedophile because someone barred men from her world. With domestic violence advocates as friends, who needs enemies.
This shelters came up with a novel approach to fixing the pedophile problem. Male children over the age of thirteen are barred from staying there. Too troublesome. The family broke up when the father was thrown out of the house. Now a second break up is happening with the teenage boys. Perhaps a relative has one bed available. Maybe the family of a high school friend would take him in their home. If neither option works then that is okay. He can move in with his father. Then they will both be sleeping in the car down by the river.
Children of these parents also suffer. They used to have their own bedroom in a safe town with good schools. First they have a shelter, then Section 8 public housing. An urban school. Maybe good-maybe not. Kids learn how to be tough in an urban environment. The kids might go bad or they could come out just fine. But there will be no clunky car as a teenager. There will be no saving fund for college. There will be no monetary gift to use as a down payment for a starter home. This tradition of the older generation giving the younger generation a financial leg up has been ruin due to the older generation's lack of money. Financially, the older generation is merely treading water. It will take generations after these present two generations to repair the economic damage to these families.
So we are stuck with funding these shelters for a while. These women and children have no place left to go. Some of you guys may think that these feminist caused the problem and then created the solution. But homeless shelters are not a solution. They are just barely a band aid.
The remaining money under VAWA goes to the United States Department of Justice for the Office of Violence Against Women (OVW). As long as OVW exists then the government is at war with men. As long as there is a pogrom against men, then women and children are going to end up as collateral damage. So there is no need for discussion about OVW going. The only thing we need to figure out is which of the two ways we can use to get rid of them-the easy way or the hard way.
And boys, do not try to burn down Washington's Dept. of Justice Building in an effort to get rid of the Office of Violence Against Women. Their offices are over at N Street.
The easy way is using Congress. The VAWA comes up for funding every five to seven years. Next time it comes up, Congress votes no and everyone at the OVW gets a pink slip in late September. Nice and simple except nothing is simple in Washington. We, the people out here in the sticks, do not always know what the dynamics are in Washington. There might be one method of getting Congress on course. Have Congress demand that the Attorney General get, and release the arrests figures. Or have the President order it. He is usually fearless after he makes up his mind. And this is too large and too well known to continue the Washington plausible deniable routine. Then they will know how much trouble they are in because of these arrests.
There are 220 million adults 18 or older in this country of both sexes. If my figure of 36 million is correct, then that is 16.4% of the adults have been arrested. It could be as high as 55 million or 25%. It might be as low as 22 million or 10%. Whatever the number there are two things that Congress should know. First, is the fellow who discovered the arrests in Minneapolis back in 1992 said do not use it because it does not work. And second, the people arrested now constitute a Fifth Column here in the United States. Our loyalty to Washington is gone. But what did these genuises on the Potomac expect? They have harmed our children. If they think Al Qaeda is a pain in the ass, wait to they see what Americans can do once their fuse is lit.
I am certain the Attorney General will sit for months on the request for the number of domestic violence arrests. Then he will explain that they do not readily have the number and that some sort of Manhattan Project effort will be needed in time and money. Nonsense. When Washington started these arrests in 1984 over 6.3 personal computers were sold here in the U.S. That figure does not include all the mini's, midi's and mainframe computers sold that year. There is no way they can pretend that this data does not exist in electronic storage. A request to Ohio for the arrests 1984-2010 would tie up a state clerk for an hour, including their 15 minute coffee break. Time for the truth boys and girls. Because this is not going away.
The hard way is more time consuming, cost more money and is full of headaches. Because the only way of removing a department from the federal government without the consent of Congress is to take out the entire federal government.
The first time I heard that, I said that is ridiculous. We cannot run this country without a federal government. But we will replace the old government with something new and improved. The new government would honor the debts incurred by the old government. There are a lot of useful reasons for starting with a clean slate.
The bipartisan debt commission released their recommendation for cleaning up the $14 trillion we have borrowed over the years. Convention wisdom has it that Congress has no stomach for any of the recommendations.
But a new government could install those recommendation on day one. Three years later, most Americans will not remember that anything is different. The old government laid off its employees when it closed. The new government is hiring. But instead of 65,000 employees at the Dept. of Education, the new government is only hiring 45,000. Instead of an average federal wage of $70,000 a year, the new average will be $52,000. The new government will have to write a tax code. Everyone pays 15% with no deductions. How many IRS employees could you get rid of if there were no more deductions? Any thing is possible with a new government.
Normally over-throwing a elected government is considered treason. Treason is punishable by death here in the United States. But there is one way of over throwing the government. That is through the ballot box. Then it is not treason but democracy. Allegedly, Washington is in favor of democracy, particularly if their candidate wins.
There is no legal mechanism in the Constitution or the Federal code of the United States for dissolving the government of the United States. So that is what we need first. Congress would need to write it. We get them to do it through the ballot initiative.
A ballot initiative is when enough registered voters sign a petition to get a question on the ballot for the next election. The following would be a sample of what the question would look like in New Hampshire.
That all elected representatives from the state of New Hampshire to both houses of the United States Congress are to propose and advance a bill that would set up a legal mechanism to dissolve the United States government should the people decide to do so in a general election by a simple majority.
If this initiative passes in all 50 states then Congress will be stuck. They will have to write the law to dissolve. If they do not I suspect within ten years they will be standing in a stairwell at the British or French embassy with a suitcase in hand waiting to get to a rooftop helicopter. I doubt if they will be thinking about the humiliation of being thrown out of the country. They will be far too busy worrying about what will happen if the mob gets their hands on them.
Washington has not got a friend in the world. Even the British and Israelis loath them now. Kind of a bad time to be losing domestic support. And what they done over the last 25 years? They have wiped out the middle class pandering to a special interest group of bigots. And in typical Washington fashion, they did not even know they did it.
This Ivy League inbreeding in Washington has produced an elite that knows what best. Everyone else-husbands, wives, police officers, prosecutors, judges, attorney generals and guardian ad litems-are to shut up and do what they are told. The rule of law is gone, replaced by the policies, procedures and protocols of The Second Set of Books. Which means the federal government will be going shortly. For the government being unable to deliver the rule of law is like an auto mechanic who claims he does not know how to change the engine oil. A certain minimum competency is required. So it looks like the parents of the Washington elite were right. One can be too smart for their own good.
Betty Friedan wrote that the feminist revolution, like any revolution, would have its excesses. Losing the rule of law is too great to call it a mere excess. It is a catastrophe. It is the heart, mind and soul between the people and their government. These feelings of betrayal by losing it may be permanent. I have 21 years of Army service going back to the Vietnam War. My loyalty to the government should be a given. It is gone. I am certain it will never return regardless of how long I might have lived.
It was another woman that lead us in to this decision to clean house inside the beltway. Something she taught us fifty years ago. You simply look at those folks in Washington and then ask yourself the old Ann Landers question, "Am I better off with them, or without them? Are my children better off with them, or without them?" They are sinking like stones.
Washington, DC was chose as the capital because it was the geographical center or the old Colonies. Today, the geographical center of the country is just west of St. Louis Missouri. The new government can set the capital anywhere in the United States it wants. Imagine how many rodents, insects and parasites they could lose by moving 1500 miles west.
Whether you replace the federal government or not, men are still going to need a legal defense center for men. Something like the NAACP used to get black people their rights. The only checks and balances in the Second Set of Books is the First Set of Books. Which means lawsuits. Now I know you guys are broke. Some of you have had your wives and kids thrown into homelessness. So I completely understand when you tell me that you are broke. But if everyone who has been arrested throws in $10.00 a year then the legal defense center will have a war chest of $360 million. You can buy a whole bunch of lawsuit with that kind of money.
The Ball family has been supplying sergeants to the Army since at least the Revolutionary War. Elijah served as a sergeant in Cushing's Regiment at the Battle of Bennington. His commanding officer was a general from NH with a name of John Stark. General Start was a clever warrior. He was responsible for the bulk of the heavy casualties the British suffer at their victory at Bunker Hill. His orderly, fighting withdrawal allowed the other units on the hill to not only retreat but collect their wounded on the way out.
General Stark would repeat this performance on three hill tops outside the village of Bennington VT one hot August day in 1777. At the end of the battle, the British lost over 900 men killed or captured. The Colonists suffered 30 dead. Two months later, the depleted British army would surrender at Saratoga. That victory at Saratoga would bring the French into the war. John Stark was the most competent general this country ever produced. For that reason alone his men loved him.
But as brilliant as he was on the battlefield, General Stark would become even more famous for something he said. In 1809 the veterans of Bennington decided to have one last reunion. A delegation called on the General with his invitation. But the General was old and frail. He could not attend. But he did send a message, "You tell the boys I said live free or die. That death is not the worst of evil." Since 1945 the State of New Hampshire has stamped Live Free or Die on every pen, coffee mug, license plate and highway sign that they have gotten their hands on.
I think the General and his sergeant would be please that his words have elevated from the novelties and bric-a-brac to something more dignified like a courthouse door. Neither of them would give a second thought to the mess left over after the fire was extinguished. War has always been a grim business. Civil wars are usually worse.
But they would be trouble by the new enemy. Oh, they understood when a government betrays it people. They took up arms against the super power of their day to get relief for their grievances. But the enemy we face now is the government that these men birthed at places like Bennington, Saratoga and Bunker Hill. Government is no different than the food in a refrigerator. Given enough time both will go bad.
The smartest person I knew in this life was my mother. Perhaps that is true of all of us. Maybe I just got lucky. She was a nurse by trade. She worked in a time when Western medicine made that final transition from butchery to science. But it would not be her nursing skills that made her extraordinary. No, it would be this one incredible knack she had that I had only modest success at mimicking in my life. If she had something important to say to you, she would say and then never mention it again. She would talk about it if you raised the issue. But she never mentioned it twice on her own. And, oddly, you always heard her.
But she did have one favorite saying. I must have heard in a thousand times in the eighteen years I lived under her roof. It always came at the end of the conversation as she peeled away to see if it was time for Perry Mason or Lawrence Welk. She would turn her head to the side, and over her shoulder she would say, "And the only thing you really have in this world is your family." Now, thanks to the United States Government, neither we nor our children have that.
. I have three things to say to my children. First, Daddy loves you. Second, you are my three most favorite people in the world. And last, that you are to stick together no matter how old you get or how far apart you live. Because it is like Grandma always said. The only thing you really have in this world is your family.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
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Creative Commons » gautam john http://creativecommons.org Share, reuse, and remix — legally. Fri, 17 May 2013 00:22:14 +0000 en-US hourly 1 http://wordpress.org/?v=3.5.1 Letter from featured superhero Gautam John of Pratham Books http://creativecommons.org/weblog/entry/24780 http://creativecommons.org/weblog/entry/24780#comments Tue, 16 Nov 2010 18:26:18 +0000 Allison Domicone http://creativecommons.org/?p=24780 I’m pleased to introduce Gautam John, one of our exceptional CC Superheroes, who will tell you in his own words why he supports Creative Commons and why you should too. Gautam John is Manager of New Projects at Pratham Books, a children’s book publisher in India that truly embodies a spirit of openness and innovation on the web. They’ve now released 105 children’s books (in English, Hindi, Tamil, Telugu, Kannada, Marathi and Gujarati) as well as loads of delightful illustrations under a CC-BY license so they can easily be shared and even remixed to create new content relevant to other languages and cultures. Here is Pratham’s story. Join Gautam in supporting Creative Commons with a donation today.
Gautam John
Donate
“As a children’s book publisher, we have always struggled to be as inclusive as we can. However, as a small non-profit, we do function under severe constraints of time, money and ability to live up to this ideal and it was the Creative Commons model of licensing that allowed us one of our biggest moments of joy — when our books were made available as Braille and Audio Books for print impaired children across the world. Without the Creative Commons licensing model and philosophy, we would not have been able to engage with multiple organizations to help build inclusion and scale.
At Pratham Books, we have a very simple mission – “A Book in Every Child’s Hand” and this drives all of our work and we constantly test what we do against this goal. The mission has two parts, one is to create more reading matter such that there is more available for children to read and the second really is a corollary – that we need to be able to get books to where children need it the most and that the books need to be culturally and linguistically relevant as well.
This is where our challenge lies – to massively scale the production of high quality, low-cost children’s books for a massively multi-lingual and multi-cultural market. Looking at this challenge it is fairly obvious that this is not a problem that any one organization can solve. The solution has to be scalable, flexible and catalyse our fundamental mission as well.
At this point, we realised that there were several internal questions to answer and some of them painfully introspective. Questions as to whether the books we create and distribute have to be a Pratham Book, whether it implied that every book must be paid for by either the reader or an intermediary and, from being a publisher, questions as to whether we are gatekeepers of content or content curators, how we could create infinite good with finite time and resources and most importantly, how we can create more value than we capture?
Having answered most of these questions using “openness” (whereby, we asked ourselves whether allowing unrestricted access to use and re-use our content furthered our mission) as a test and finding that it did fit our mission, the second set of questions to answer was more technical – how, as a small non-profit, do we do this and not find ourselves overwhelmed. It was at this point that we had a moment of realization – that reading is an extremely social activity and that there are communities and organizations who were more than ready to help us achieve our goals.
It was at this juncture that we hit upon the Creative Commons licensing model as one that would help us achieve many of our aims of flexibility, scalability and being able to help catalyse our mission of a book in every child’s hands. In particular, three things stood out – a shared value system of sharing and openness, a community that was deeply embedded in these ideals and, from our perspective, it was scalable because it allowed us to license content to multiple organizations and individuals, both known and unknown, with a one time effort of releasing them under a Creative Commons license as opposed to the traditional model which involves time consuming negotiations and discussions with each known organization or individual who wants to use our content.
As an organization, we did spend some time choosing a license and, from our perspective, a choice between openness and sharing which reduced to a choice between the Attribution and Attribution-Share-Alike license. We have decided that the Attribution license will be our default license with a fall-back to the Attribution-Share-Alike license in cases where needed. It is best said by P2PU “it emerged that our choice lay between two licences: Creative Commons Attribution and Creative Commons Attribution Share Alike …chose to use Creative Commons licences because Creative Commons have become a global standard and are supported by a large international community. Both licences are Free Culture licences and are more permissive than any of the other Creative Commons licences. In other words, the choice was not between two extremes but between two open licences at the same end of the licence spectrum.” Given that our goal was being as open as possible, it followed that our license choices were essentially around licenses that allowed for the greatest possible use and re-use because our initial hypothesis was, and continues to be, that being open allows us to fulfill our mission better than a traditional copyright model allows.
We now use Creative Commons licenses everywhere! We license entire books under CC-BY and CC-BY-SA licenses, we license our illustrations similarly and even photographs and other publicity material too. Over the last year we have been building the foundations for a social publishing model – where we curate communities that are passionate about reading and help us create content. Such a model rests on the idea of a participatory culture and an essential ingredient is a permissive licensing strategy – Creative Commons licenses offers us this, a large community with shared values and an ecosystem to tap in to.
While this licensing and publishing model works well in theory, it has been extremely heartening for us to see it come to life – our communities have created multiple derivative works ranging from iPad and iPhone applications, to porting our works to OLPC laptops, to creating entirely new books from existing illustrations and, my personal favourite, creating versions of our books for the print impaired – from DAISY and Braille books to rich audio books such that our mission truly does encompass every single child.
I firmly believe that we would not have been able to achieve what success we have had without the help of Creative Commons licensing. These licenses and the values that they stand for are vital to building and strengthening a digital commons from which we all benefit. I hope you will consider supporting Creative Commons and licensing content that you own or control such that we all benefit from the growth of the commons.”
Follow Gautam on Twitter.
Special thanks to Maya Hemant from Pratham Books for getting all content (books, images) up online and for managing the Pratham community.
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Error!
Success!
VisualStudio.NET vNext Feature Request: Multi-target IntelliSense
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kicks
VisualStudio.NET vNext Feature Request: Multi-target IntelliSense (Unpublished)
With all the partitioning happening in the WPF-like framework world (WPF, Silverlight, Silverlight for Windows Phone 7) there's a need for assistance from the tools in creation of code that works in all (several) of them. Here is an idea for one such feature for Visual Studio.
Kicked By:
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Minireview
Tetraodon genome confirms Takifugu findings: most fish are ancient polyploids
Yves Van de Peer
Author Affiliations
Bioinformatics and Evolutionary Genomics, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Ghent University, Technologiepark 927, B-9052 Ghent, Belgium
Genome Biology 2004, 5:250 doi:10.1186/gb-2004-5-12-250
The electronic version of this article is the complete one and can be found online at: http://genomebiology.com/2004/5/12/250
Published:25 November 2004
© 2004 BioMed Central Ltd
Abstract
An evolutionary hypothesis suggested by studies of the genome of the tiger pufferfish Takifugu rubripes has now been confirmed by comparison with the genome of a close relative, the spotted green pufferfish Tetraodon nigroviridis. Ray-finned fish underwent a whole-genome duplication some 350 million years ago that might explain their evolutionary success.
Minireview
In 1993, Sydney Brenner and colleagues [1] proposed sequencing the pufferfish genome as a cost-effective way to identify and characterize human genes. The genome of the pufferfish is only about one-eighth of the size of that of human but was expected to contain a similar gene repertoire. Ten years later, not only has a draft genome sequence been released for Takifugu rubripes (Fugu, also known as the Japanese or tiger pufferfish) [2], but also for Tetraodon nigroviridis (green spotted pufferfish) [3], a close relative that diverged from Takifugu 18-30 million years ago (Mya). By comparing the two pufferfish genomes with that of human, several hundred novel human genes have already been uncovered, as was predicted by Brenner and colleagues [1]. But the pufferfish genome sequencing projects have also yielded a surprising finding: ray-finned fish (Actinopterygii), such as pufferfish might have more genes than lobe-finned fish (coelacanths and lungfish) and land vertebrates, because of additional gene-duplication events [4]. The recent release of the Tetraodon genome sequence [3] provides overwhelming evidence that a genome-duplication event did indeed occur early in the evolution of ray-finned fish.
A fish-specific genome duplication
Some of the first data pointing to a possible genome duplication in fish came from Hox genes and Hox gene clusters. Hox genes encode DNA-binding proteins that specify cell fate along the anterior-posterior axis of bilaterian animal embryos and occur in one or more clusters of up to 13 genes. Whereas lobe-finned fish, amphibians, reptiles, birds and mammals have four clusters, extra Hox gene clusters have been discovered in zebrafish, Medaka, Nile tilapia and pufferfish [4]. The observation that such distantly related species [5] all have seven or eight Hox gene clusters suggested the occurrence of an additional genome-duplication event in the ray-finned fish lineage before the divergence of most teleost (bony fish) species. More recent comparative genomic studies have turned up many more genes and gene clusters for which there are two copies in fish but one in other vertebrates [6]. The findings that different paralogous pairs seem to have originated at about the same time, that different fish species seem to share ancient gene duplications, and that different paralogs are found on different linkage groups in the same order as other duplicated genes, all support the hypothesis that these genes arose through a large-scale gene-duplication event. It is worth noting, however, that some authors have argued that an ancestral whole-genome-duplication event was not responsible for the abundance of duplicated fish genes [7].
Additional evidence for a genome duplication in ray-finned fish was provided by analyzing the complete Takifugu genome, a draft sequence of which was published in 2002 [2]. Two recent studies identified duplicated genes in this genome and used phylogenetic trees to estimate the ages of these duplicates [8,9]. Vandepoele et al. [8] constructed phylogenetic trees for all gene families containing between two and ten duplicated Takifugu genes, which amounts to a total of 3,077 families. For each gene family, the relative date of duplication events was determined to test whether gene duplications occurred before or after the split between fish and land vertebrates. To this end, neighbor-joining trees were created for each of the Takifugu gene families with homologous sequences from mouse and human. Absolute dating of duplication events was achieved through inference from linearized trees [10]. In such trees - where branch length is directly proportional to time - the split between ray-finned fish and land vertebrates, dated at 450 Mya, was used as a calibration point for the dating of gene-duplication events. A major fraction (about one-third) of the duplicated genes in Takifugu could be ascribed to a large-scale gene-duplication event specific to the fish lineage, which was estimated to have occurred about 320 Mya (Figure 1).
Figure 1. A phylogenetic tree showing the vertebrate phylogenetic relationships and superimposed pufferfish gene-duplication events. (a) A generally accepted tree illustrating the relationships between several vertebrate species. The gray horizontal bar denotes the fish-specific genome-duplication event inferred from absolute dating of Takifugu paralogs. The broken line indicates the position of the duplicated copy of the Takifugu genome that originated between the divergence of gar and the bony tongues. (b) The bar chart shows the number of paralogous genes that could be dated through the construction of linearized trees. Modified from [8,9].
A very similar approach was followed for the analysis of the Takifugu genome by Christoffels et al. [9], who obtained essentially the same result: by constructing linearized trees, the whole-genome-duplication event was estimated to have occurred approximately 350 Mya. To test whether the sudden increase in the number of duplicated genes in the Takifugu genome was the result of an entire-genome duplication rather than an increased rate of independent tandem-duplication events, both Vandepoele et al. [8] and Christoffels et al. [9] investigated the appearance of duplicated genes in duplicated blocks on chromosomes. Statistically significant regions of micro-colinearity were identified within the complete Takifugu genome, showing the same gene content and gene order. Indeed, both studies reported a large number of duplicated genes in so-called paralogons - homologous genomic segments that can be proved to have been created by duplication [11] - and concluded that most findings were congruent with a large-scale, probably whole-genome duplication event in a ray-finned ancestor that gave rise to the Takifugu and other fish lineages.
Comparing genomes
Because of the highly fragmented nature of the initial Takifugu genome assembly, it was difficult to prove that the large-scale gene-duplication event had indeed affected the whole genome. The recent release of the well-assembled Tetraodon genome [3] seems to have settled this issue in two ways. First, Jaillon et al. [3] analyzed the chromosomal distribution of ancient duplicates and observed that genes on one chromosome have a strong tendency to have duplicate copies on a single other chromosome. As would be expected from a whole-genome-duplication event, all chromosomes are involved. Second, by using a comparative approach in which they compared the Tetraodon genome with that of human, which has not undergone the genome-duplication event (Figure 1), Jaillon et al. [3] showed that almost every region in the human genome clearly corresponds to two regions in the Tetraodon genome. This type of comparative analysis (Figure 2) has proved very powerful for unveiling genome-duplication events. Recently, such an approach provided overwhelming evidence for the long-suggested [12], but contested (see, for example, [13]) ancient whole-genome duplication in the yeast Saccharomyces cerevisiae, by comparing its genome with that of different relatives that diverged prior to the duplication events [14,15].
Figure 2. Uncovering genome duplications through comparative analysis with related sequences. The hypothetical genomes of two related organisms are shown, each containing the same set of genes. Both genomes are initially identical, but the genome of Organism 1 is duplicated, resulting in a second identical set of chromosomes and genes. After some time, homologous chromosomes lose a different set of genes, keeping two copies for only a minority of the duplicated genes. For the sake of simplicity, the genome of Organism 2 is assumed to remain unchanged. Within Organism 1, the only evidence for a duplication event comes from the conserved order of the anchor points formed by genes 1 and 11 (indicated by boxed regions). Comparison with the genome of Organism 2, however, shows a pattern of so-called 'double conserved synteny' where the duplicated nature of Organism 1 is revealed.
A comparative analysis between the human and Tetraodon genomes has also allowed inference of the basic structure of the ancestral bony vertebrate genome, and the reconstruction of much of the evolutionary history of ancient and recent chromosomal rearrangements leading to the modern human karyotype. By matching up the genes on the Tetraodon chromosomes with homologs on human chromosomes, Jaillon et al. [3] inferred that the ancestor of both fish and land vertebrates had no more than 12 chromosomes, a number that has been previously suggested on the basis of linkage relationships between zebrafish, Medaka, and human [16]. Comparison of the genomes of Tetraodon and human also showed that chromosome evolution in both lineages differed considerably. Whereas all but one of the ancestral Tetraodon chromosomes had not undergone interchromosomal exchange for 450 Mya, only one human chromosome was similarly undisturbed. A possible explanation for the difference in genome evolution might be the massive integration of transposable elements in the human genome, with an increased overall frequency of chromosome breaks as a result [3].
Evolutionary implications
As mentioned above, on the basis of previous analyses of the Takifugu genome, the whole-genome-duplication event in fish is thought to have occurred somewhere between 300 and 350 Mya [8,9]. An interesting question is whether this date correlates with a decisive period in the evolution of the fish. For instance, if the genome duplication had been responsible for the biological diversification and large number of ray-finned fish, as suggested previously [4,17], it must have occurred prior to the radiation of most fish lineages. The class Actinopterygii includes more than 23,500 species [18], of which the vast majority are teleosts or ray-finned fish. Interestingly, all older, more basal groups of ray-finned fish, namely Polypteriformes (bichirs), Acipenseriformes (sturgeons and paddlefish), Semionotiformes (gars), and Amiformes (bowfin), have only a few extant species (Figure 1). Most members of these basal actinopterygian lineages are considered to be 'living fossils', because their morphology has remained unchanged over very long evolutionary time periods.
In a recent study, Hoegg et al. [19] have tried to determine the timing of the duplication event in relation to the origin of lineages of teleost and 'nonteleost' fish by sequencing three nuclear genes - fzd8, sox11 and tyrosinase - from sturgeons, gars, bony tongues, and a tenpounder. For these three genes, two copies have been described previously in derived teleost model species, such as zebrafish and pufferfish, but only one orthologous copy has been found in tetrapods. The specific clustering of the genes in individual gene trees for these three genes and a dataset of concatenated genes support the hypothesis that the fish-specific genome-duplication event took place after the split of the Acipenseriformes and the Semionotiformes from the lineage leading to teleost fish, but before the divergence of Osteoglossiformes (bony tongues) and the other more derived groups of fish (Figure 1). This is in good agreement with the recent analyses of the Takifugu genome, as fossil data age the Semionotiformes at between 245 and 286 million years, whereas molecular estimates for the Amiiformes, which are of approximately the same age as the Semionotiformes, hint at a separation from the Teleostei stem lineage about 367-404 Mya. Likewise, molecular data suggest an age of 335 million years for the Osteoglossiformes [19]. The inferred relative and absolute dates for the fish-specific genome duplication event seem to separate the species-poor branching lineages from the species-rich teleost lineages, providing evidence that the fish-specific genome duplication might be related causally to an increase in species and morphological diversity.
On the basis of isozyme studies, Werth and Windham [20] developed a model in which the 'reciprocal silencing' of genes in geographically separated populations would promote speciation. A few years ago, this idea was revived in a model called 'divergent resolution', in which the loss or silencing of gene duplicates was postulated to be more important for the evolution of species diversity than the acquisition of new functions by duplicated genes. Divergent resolution occurs when different copies of a duplicated gene are lost on different chromosomes in different populations, thereby creating genetic barriers for reproduction between them [21,22]. Divergent resolution and lineage-specific subfunction partitioning [17] can promote incompatibility among populations within a species, and thus might facilitate evolutionary radiation. Gene duplications might, therefore, bring about rapid speciation in populations fixed for different copies of a duplicated locus. The fish-specific genome duplication has created many duplicates that could be divergently resolved. Potentially, such genes have played a prominent role in the radiation of the teleosts. Further studies of the genes encoded in these fish genomes may shed light on how important the fish-specific whole-genome duplication has been in the evolution of the ray-finned fish.
References
1. Brenner S, Elgar G, Sandford R, Macrae A, Venkatesh B, Aparicio S: Characterization of the pufferfish (Fugu) genome as a compact model vertebrate genome.
Nature 1993, 366:265-268. PubMed Abstract | Publisher Full Text
2. Aparicio S, Chapman J, Stupka E, Putnam N, Chia JM, Dehal P, Christoffels A, Rash S, Hoon S, Smit A, et al.: Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes.
Science 2002, 297:1301-1310. PubMed Abstract | Publisher Full Text
3. Jaillon O, Aury J-M, Brinet F, Petit J-L, Stange-Thomann N, Mauceli E, Bouneau L, Fischer C, Ozoef-Costaz C, Bernot A, et al.: Genome duplication in the teleost fish Tetraodon nigroviridis reveals the early vertebrate proto-karyotype.
Nature 2004, 431:946-957. PubMed Abstract | Publisher Full Text
4. Meyer A, Schartl M: Gene and genome duplications in verterbrates: the one-to-four (-to-eight in fish) rule and the evolution of novel gene functions.
Curr Opin Cell Biol 1999, 11:699-704. PubMed Abstract | Publisher Full Text
5. Chen W-J, Orti G, Meyer A: Novel evolutionary relationships among four fish model systems.
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Research
Long noncoding RNA genes: conservation of sequence and brain expression among diverse amniotes
Rebecca A Chodroff1,2, Leo Goodstadt3, Tamara M Sirey1, Peter L Oliver3, Kay E Davies1,3, Eric D Green2, Zoltán Molnár1* and Chris P Ponting1,3*
Author Affiliations
1 Department of Physiology, Anatomy, and Genetics, Le Gros Clark Building South Parks Road, University of Oxford, Oxford OX1 3QX, UK
2 Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, 50 South Drive, Building 50, Room 5222, Bethesda, MD 20892, USA
3 MRC Functional Genomics Unit, Le Gros Clark Building, South Parks Road, University of Oxford, Oxford OX1 3QX, UK
For all author emails, please log on.
Genome Biology 2010, 11:R72 doi:10.1186/gb-2010-11-7-r72
The electronic version of this article is the complete one and can be found online at: http://genomebiology.com/2010/11/7/R72
Received:4 March 2010
Revisions received:17 May 2010
Accepted:12 July 2010
Published:12 July 2010
© 2010 Chodroff et al.; licensee BioMed Central Ltd.
This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background
Long considered to be the building block of life, it is now apparent that protein is only one of many functional products generated by the eukaryotic genome. Indeed, more of the human genome is transcribed into noncoding sequence than into protein-coding sequence. Nevertheless, whilst we have developed a deep understanding of the relationships between evolutionary constraint and function for protein-coding sequence, little is known about these relationships for non-coding transcribed sequence. This dearth of information is partially attributable to a lack of established non-protein-coding RNA (ncRNA) orthologs among birds and mammals within sequence and expression databases.
Results
Here, we performed a multi-disciplinary study of four highly conserved and brain-expressed transcripts selected from a list of mouse long intergenic noncoding RNA (lncRNA) loci that generally show pronounced evolutionary constraint within their putative promoter regions and across exon-intron boundaries. We identify some of the first lncRNA orthologs present in birds (chicken), marsupial (opossum), and eutherian mammals (mouse), and investigate whether they exhibit conservation of brain expression. In contrast to conventional protein-coding genes, the sequences, transcriptional start sites, exon structures, and lengths for these non-coding genes are all highly variable.
Conclusions
The biological relevance of lncRNAs would be highly questionable if they were limited to closely related phyla. Instead, their preservation across diverse amniotes, their apparent conservation in exon structure, and similarities in their pattern of brain expression during embryonic and early postnatal stages together indicate that these are functional RNA molecules, of which some have roles in vertebrate brain development.
Background
Whilst only approximately 1.06% of the human genome appears to encode protein [1,2] at least four times this amount is transcribed into stable non-protein-coding RNA (ncRNA) transcripts [3-5]. Unfortunately, the biological relevance of the vast majority of this extensive and interleaving network of coding RNAs and ncRNAs remains far from clear. One possibility is that many ncRNAs result simply from transcriptional 'noise'. If so, their sequence and transcription might be expected not to be conserved outside of restricted phyletic lineages. Indeed, the finding that only 14% of the well-defined mouse long intergenic ncRNAs (lncRNAs) identified in the FANTOM projects [6,7] have a transcribed ortholog in human (based on analyses of known EST and cDNA data sets) [2] argues against their functionality. Similarly, known human intergenic lncRNA loci are generally not conserved in sequence at statistically significant levels in the mouse genome [3,8,9], and there is little evidence for conserved expression of intergenic regions (including lncRNAs) between mouse and human [10].
On the other hand, our preconceptions of lncRNA functionality might be greatly prejudiced by our long-standing knowledge of protein evolution. Just because functional protein-coding sequence is highly constrained, this need not necessarily imply that largely unconstrained non-protein-coding sequence, free from the need of maintaining an ORF and producing a thermodynamically stable protein product, is not functional. Indeed, even well-known examples of functional mammalian lncRNAs, such as Gomafu [11], Evf-2 [12], XIST [13], Air [14], and HOTAIR [9], exhibit poor sequence conservation across species. Moreover, there is evidence for significant, albeit modest, evolutionary constraint within lncRNA loci compared to neutrally evolving DNA [15-18]. In addition, as with mRNAs, many lncRNAs are subject to splicing, polyadenylation, and other post-transcriptional modifications, and their loci tend to be associated with particular chromatin marks [15]. However, whether the observed chromatin marks and purifying selection are most frequently directed towards the transcribed lncRNA, the process of transcription, or the underlying DNA sequence remains unknown [19-21].
In support of functional roles for lncRNA loci, many lncRNAs have been shown to be developmentally regulated and/or expressed in specific tissues. For example, a computational analysis of in situ hybridization data from the Allen Brain Atlas identified 849 lncRNAs (out of 1,328 examined) showing specific expression patterns in adult mouse brain [22]. Similarly, 945 lncRNAs were found to be expressed above background levels in a microarray screen of mouse embryonic stem cells at various stages of differentiation [23]. A follow-up study found that 5% of approximately 3,600 analyzed lncRNAs are differentially expressed in forebrain-derived mouse neural stem cells subjected to various developmental paradigms [24]. Such regulated expression patterns can perhaps be attributed to lncRNA loci tending to cluster near brain-expressed protein-coding genes and transcription factor-encoding genes associated with development [15,17,25].
Nevertheless, it is important to stress that the above-mentioned studies focused on only one species, namely the laboratory mouse. There is a clear and substantial need to investigate the evolution and expression of specific lncRNA loci for more diverse species, for example birds, whose lineage separated from that of mammals approximately 310 million years ago [26]. However, few, if any, studies have identified orthologous lncRNAs shared between birds and mammals, let alone investigated either their expression in homologous developmental fields or adult anatomical structures, or their molecular functions. Whilst one study found that Sox2ot is both dynamically regulated and transcribed from highly conserved elements in chicken and zebrafish [27], this locus overlaps with a protein-coding gene (Sox2), a pluripotency regulator, and thus is not intergenic. A more comprehensive study of full-length chicken cDNA sequences identified 30 transcripts that could be aligned with RIKEN-identified mouse lncRNAs, although their expression in developing chick embryos was undetectable [28]. Even Xist, which is involved in chromosome-wide × inactivation in eutherians, is not conserved as a lncRNA in birds, as its avian ortholog is protein-coding [29].
In this study, we used a multi-disciplinary approach to investigate a select group of highly conserved lncRNAs that are expressed within the embryonic and early postnatal mouse brain. We report the characterization of four such lncRNAs, demonstrating that they are expressed at experimentally detectable levels, are tissue-specific and developmentally regulated, and are conserved in transcript structure and expression pattern across diverse amniotes during brain development. To our knowledge, this is the first description and investigation of lncRNA loci with orthologs present in eutheria, metatheria (marsupials), and birds. As these lncRNAs do not differ substantially from protein-coding genes in their sequence or expression properties, we propose that they are novel RNA genes that are likely to confer important functions among these diverse amniotes. Our observations provide the first indications that investigation of lncRNA orthologs in amniote model organisms will be informative about their contributions to human biology.
Results
lncRNA selection
We started with a set of 3,122 well-characterized intergenic lncRNAs derived from FANTOM 2 and 3 consortia collections of full-length noncoding transcripts in the mouse [6,7,18]. While transcripts with evidence of protein-coding capacity had already been discarded, we removed additional lncRNAs that overlap either with more-recently annotated mouse protein-coding genes or with alignable protein-coding genes from other species. We also discarded lncRNAs transcribed in close proximity (<5 kb) of annotated protein-coding genes in order to reduce the chances of inadvertently considering untranslated regions or alternative transcripts of these genes. Of the remaining set of 2,055 lncRNA transcripts, 1,209 (59%) harbor strongly constrained sequence, based on overlap with phastCons-predicted conserved elements (Figure 1b) [30], consistent with a recent report [16]. On average, 10.6% and 10.9% of the lncRNA sequences (including and excluding introns, respectively) overlap phastCons-predicted conserved elements.
Figure 1. Sequence conservation among lncRNAs. (a) Conservation across a generic lncRNA locus, based on 877 mouse multi-exon lncRNAs. We sampled 200 evenly spaced bases across each region listed, with regions containing fewer than 200 bases sampled entirely. The graph shows the average vertebrate phastCons score at each genomic position across all multi-exon lncRNA loci. Note phastCons score peaks within the putative promoter region (200 bp upstream) and near donor and acceptor splice sites (analysis inspired by Figure 25a in [31]). (b) Overlap between vertebrate phastCons-predicted conserved elements and mouse lncRNA exons. Of 2,055 lncRNAs with signatures of purifying selection initially identified in mouse [18], 1,095 contain exons that overlap phastCons-predicted vertebrate conserved elements (log-odds score range 1 to 1,000) [30]. Depicted is a histogram showing the percentage of each lncRNA transcript that overlaps a phastCons-predicted vertebrate conserved element. The relative positions of three selected lncRNAs (AK082072, AK043754, and AK082467 with overlaps of 36.7, 44.8, and 51.7%, respectively) are shown.
To compare the evolution of lncRNA loci with protein-coding gene evolution, we next constructed a generic locus from 877 multi-exon lncRNA loci, and annotated it according to the presence of conserved sequence elements (Figure 1a). A similar portrait of evolutionary conservation for protein-coding genes was presented by the Mouse Genome Sequencing Consortium (Figure 25a in [31]). As seen for protein-coding genes, sequence conservation is not uniformly distributed across various features (exons, introns, and upstream and downstream regions) of a generic multi-exon lncRNA locus (Figure 1a). The putative core promoter region (here defined as 200 bp upstream of each lncRNA transcription start site (TSS)) is generally under greater evolutionary constraint than lncRNA exonic sequence, in agreement with previous reports [6,16,18]. Constraint peaks at 0.19 (range between 0 and 1), 43 bp upstream of the normalized TSS, as previously observed for human and mouse promoter sequence [32]. Just as for protein-coding genes [31], the generic lncRNA locus' first, middle and last exons tend to be under greater evolutionary constraint than its introns, with average phastCons scores peaking in close proximity to splice sites.
To establish whether lncRNAs are conserved in expression as well as in sequence, we sought to select a small number of mouse lncRNAs and investigate their putative orthologs in other amniotes, namely the marsupial opossum (Monodelphis domestica) and the chicken (Gallus gallus). We chose lncRNAs that are highly conserved, developmentally regulated, and brain-expressed. These criteria were used because our previous study [17] found that constrained lncRNAs with significantly suppressed human-mouse nucleotide substitution rates tended to be expressed in the mouse brain and, when developmentally expressed, to be transcribed near protein-coding genes involved in transcriptional regulation.
Accordingly, we selected three lncRNAs, each having extensive overlap with phastCons-predicted conserved elements (Figure 1b) and each expressed in embryonic or neonatal brain based on the origin of the cDNA library from which they were identified. Here, we refer to these three lncRNAs and their genomic loci according to their database accession numbers: AK082072, AK082467, and AK043754.
Structure of selected lncRNA loci
The three selected lncRNA loci harbor elements that are more usually associated with protein-coding genes. These include GT-AG donor-acceptor splice sites, polyadenylation signals, and chromatin marks in their putative promoter regions (Figures 2b,c, 3b,c and 4b,c; Figure S1 in Additional file 1). Aceview annotations [33] indicate an unspliced (single exon) transcript and single promoter for the AK043754 locus (spanning 1.75 kb on mouse chromosome 6qG1), a single canonical GT-AG intron and promoter for the AK082072 locus (39.7 kb on mouse chromosome 13qC3), and 31 different GT-AG introns in at least 16 different mRNA splice variants and 6 probable alternative promoters for the AK082467 locus (94 kb on mouse chromosome 10qC2). Each lncRNA sequence is supported by several GenBank cDNA records, representing cDNAs derived primarily from mouse embryonic or neonatal central nervous system tissues, including hypothalamus, diencephalon, cortex, cerebellum, and spinal cord. Many of the supporting GenBank records additionally support poly(A) and 5' cap structures, indicating that each lncRNA is most likely transcribed by RNA polymerase II. Chromatin marks from either mouse embryonic stem cells or adult mouse whole brain [34] are present at each putative lncRNA promoter (Figures 2b, 3b and 4b).
Figure 2. Evolutionary constraint of AK043754. (a) The genomic region of mouse chromosome 6 (chr6) encompassing the lncRNA locus AK043754 (1.7 kb) is depicted. Note the locations of flanking protein-coding genes: Grin2B (glutamate receptor, ionotropic, NMDA2B (N-methyl-D-aspartic acid)) and Emp1 (epithelial membrane protein 1). Also shown are the positions of mouse-chicken ECRs (evolutionarily conserved regions at least 100 bp in size with 70% sequence identity between the mouse and chicken genomes); ECRs within protein-coding regions are shown in blue. (b) A more detailed representation of AK043754 (single exon highlighted in orange) and its immediate flanking regions, including the 3' end of Grin2B. Below the gene structures are the positions of H3K4me1 chromatin marks (green) detected in mouse embryonic stem cells (obtained from UCSC Genome Browser), EvoFold predictions of RNA secondary structures (grey), a SinicView conservation plot [68] based on a 21-vertebrate multispecies sequence alignment (using Threaded Blockset Aligner) generated with mouse as the reference sequence, and Gmaj [66] views of alignments between mouse and the indicated species' sequences (note the detected homology with the orthologous lizard and chicken, but not frog, sequences). (c) Conservation and relative sizes of AK043754 orthologs in various species. The TSSs (arrows) and transcript lengths are depicted in each case. Note the conserved position of a polyA signal (red) and increased sequence conservation (relative to the mouse sequence) towards the 3' end. ECR, evolutionarily conserved region.
Figure 3. Evolutionary constraint of AK082072. (a) The genomic region of mouse chromosome 13 (chr13) encompassing lncRNA AK082072 (523 bp) is depicted. Note the locations of the flanking protein-coding genes: Tmem161b (transmembrane protein 161b) and Mef2C (myocyte enhancer factor 2C). (b) A more detailed representation of AK082072 (exons highlighted in orange) and its immediate flanking regions. Below the gene structures are the positions of H3K4me3 chromatin marks (green) detected in mouse brain, VISTA conserved non-coding midbrain enhancer element 268 (obtained from the UCSC Genome Browser), and a BLAT alignment of the chicken AK082072 ortholog, as well as similar tracks as those in Figure 2b. Note the detected homology with orthologous frog sequence in exon 1. (c) Conservation and relative sizes of AK082072 orthologs in various species. Note the sequence conservation (relative to the mouse sequence) at both the 5' and 3' ends and the conserved position of splice sites (green). Unlike the other vertebrate genomes considered, the zebra finch genome did not align to the proximal promoter or first exon of mouse AK082072. This apparent lack of sequence identity might reflect either an unannotated gap in its genome assembly or rapidly evolving sequence within its orthologous genomic region. Other details are provided in the legend to Figure 2. ECR, evolutionarily conserved region.
Figure 4. Evolutionary constraint of AK082467 and Rmst. (a) The genomic region of mouse chromosome 10 (chr 10) encompassing lncRNAs AK082467 (2.7 kb) and Rmst (2.7 kb) is depicted. Note the presence of the protein-coding gene Nedd1 (neural precursor expressed developmentally down-regulated protein 1) upstream of AK082467 and Rmst. (b) A more detailed representation of AK082467 and Rmst (exons highlighted in yellow and orange, respectively), microRNAs mir-1251 and mir-135a-2, and their immediate flanking regions. Below the gene structures are the positions of H3K4me3 (green) and H3K27me3 (red) chromatin marks detected in mouse brain (obtained from the UCSC Genome Browser) as well as similar tracks as those in Figure 2b. Note the detected homology with orthologous frog sequence in Rmst exons 1, 2, 4, and 11. (c) Conservation and relative sizes of AK082467 and Rmst orthologs in various species. Note the conserved splice sites (green bars) in mouse Rmst exons 1, 4, and 11 as well as the sequence conservation (relative to mouse sequence) in exons 1 and 11, but differences in total exon number among species. The 3' ends of opossum and chicken orthologs have not been experimentally verified. Other details are provided in the legend to Figure 2. ECR, evolutionarily conserved region.
Additional file 1. Figure S1: splice-site and poly(A)-signal conservation among AK043754, AK082072, and AK082467 orthologs. Figure S2: sense probe controls for in situ hybridization.
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In contrast to most protein-coding genes, the lncRNA loci each harbor at least one Evofold-predicted RNA secondary structure (Figures 2b, 3b and 4b) [35]. This reflects the general tendency of conserved brain-expressed lncRNA loci to contain such structures [17]. The three lncRNA transcripts each lack long (>100 amino acids) ORFs. While it remains possible that the lncRNAs encode short peptides, there is no evidence for constraint on their protein-coding capacity, as the frequencies of synonymous and non-synonymous substitutions across eutherians are roughly equal (that is, dN/dS ≈ 1 ± 0.16) for the longest predicted ORF of each lncRNA [36].
These findings imply that the three selected transcripts might be functional noncoding RNA genes. AK082467 is an alternative splice variant that contains the first three exons and retains the second intron of a previously described long noncoding RNA, Rmst (rhabdomyosarcoma 2 associated transcript, also known as NCRMS); the human RMST ortholog was initially identified as a differentially expressed transcript in alveolar versus embryonic rhabdomyosarcoma (a malignant soft tumor tissue), but its function remains undocumented [37]. To our knowledge, AK043754 and AK082072 have not been experimentally investigated. To examine their potential functions, we first studied the expression patterns of the three lncRNAs during mouse development.
Expression of selected lncRNAs in mouse
Analysis of the three selected lncRNAs by in situ hybridization of mouse tissues at different developmental time points revealed that each exhibits a specific expression pattern that, in general, is restricted to the brain. Our findings further suggest their expression is tightly regulated, as opposed to stochastic background transcription.
AK043754 is initially expressed in the primordial plexiform layer or preplate. This is the first of the developmental cell layers to appear during mammalian embryogenesis and is, most likely, homologous to the simpler amphibian and avian cortical structures (Figure 5a(i,ii,iv,v)) [38]. At embryonic day 17 (E17), AK043754 is expressed prominently within the marginal zone along the pial surface in a pattern similar to that of reelin-expressing Cajal-Retzius cells. Of note, the expressed transcript is also present within the ventricular zone of the ganglionic eminence, a source of GABAergic migratory neurons (including some Cajal-Retzius cells) that ultimately colonize the marginal zone, intermediate zone, and subplate; this suggests that AK043754-expressing cells might originate in the ganglionic eminence and then migrate to the preplate and marginal zone [39]. Reinforcing this transcript's potential association with inhibitory GABAergic neurons, hybridization is also seen in the latero-caudal migratory path of interneurons from the basal telencephalon to the striatum. This is best illustrated at stage E17 and within the internal granule cell layers of the olfactory bulb at postnatal day 3 (P3; Figure 5a(vii)).
Figure 5. lncRNAs are specifically expressed and developmentally regulated in the mouse brain. (a-c) Digoxigenin-labeled riboprobes complementary to AK043754 (a), AK082072 (b), and AK082467 (c) were hybridized to sagittal sections of C57BL/6J mouse brains at different development stages (E9, E13, E17, and P3). (a) The AK043754 probe hybridized to the first generated cell layer of the preplate or primordial plexiform zone (red arrowheads) at E13 (i, iv) and E17 (ii, v), the ventricular zone of the medial and lateral ganglionic eminences (black arrowhead) at E13, the latero-caudal migratory path from the basal telencephalon to the striatum (green arrowhead) at E17 (ii, v), and the hippocampus (iii, vi) and the olfactory bulb (iii, vii) at P3. Scale bar (shown in (i)) is 500 μm in (i), 543 μm in (ii), 322 μm in (iii), 292 μm in (iv), 300 μm in (v), 167 μm in (vi), and 214 μm in (vii). (b) The AK082072 probe hybridized to the hem of the embryonic cerebral cortex (blue arrowheads) and the roof of the midbrain (black arrowheads) at E13 (i, iv) and E17 (ii, v), and to the hippocampus (iii, vi), rostral migratory stream (iii, vi), and internal plexiform and granule cell layer of the olfactory bulb (iii, vi) at P3. Scale bar (shown in (i)) is 500 μm in (i), 595 μm in (ii), 422 μm in (iii), 357 μm in (iv), 386 μm in (v), and 311 μm in (vi). (c) The AK082467 probe hybridized to the optic stalk (black arrowheads) at E9 (i, v), the cortical hem (blue arrowheads) at E13 (ii, vi) and E17 (ii, vii), and the accessory olfactory bulb (iii, viii) at P3. Scale bar (shown in i)) is 500 μm in (i), 637 μm in (ii), 684 μm in (iii), 522 μm in (iv), 182 μm in (v), 177 μm in (vi), 176 μm in (vii), and 110 μm in (viii).
Cells expressing AK082072 at stage E13 primarily populate the roof of the midbrain and the cortical hem (the most caudomedial edge of the telencephalic neuroepithelium), one of the major patterning centers of the developing telencephalon and, as recently shown by Monuki and Tole and colleagues, a hippocampal precursor (Figure 5b(i,iv)) [40,41]. By stage E17, expression continues to be apparent within the roof of the midbrain, and, as illustrated at higher magnification, is strongest in the soma and outward projections of cells lining the midbrain ventricle (Figure 5b(v)). Also visible in the E17 image is the expression of AK082072 along the caudal ganglionic eminence, a major source of GABAergic neurons that preferentially migrate caudally to the caudal cortex and hippocampus [42]. At postnatal stages, AK082072 expression is restricted to the hippocampus (mostly within CA1), the rostral migratory stream, and the internal plexiform and granule cell layer of the olfactory bulb. Reinforcing our observations, a previous independent study that utilized a probe designed from another region of the AK082072 transcript yielded similar results [43].
AK082467 is expressed early in mouse brain development, with its transcription mostly attenuated after birth. The antisense riboprobe designed to an intron-spanning region of this lncRNA transcript partially overlaps the 5' region of Rmst, such that all observations could reflect the expression pattern(s) of one or both of these transcripts. Consistent with the expression pattern of Rmst described by Bouchard et al. [44], our riboprobe hybridized to the mid-hindbrain organizer region in developing mouse embryos, most clearly illustrated in Figure 5c(ii). We also found expression in two additional Pax2-expressing regions, including the optic stalk at stage E9 and within the accessory olfactory bulb postnatally (Figure 5c(i,iv)).
lncRNA orthologs in other vertebrates
AK082072, AK082467, Rmst, and AK043754 are each transcribed from regions of the mouse genome whose sequence aligns to vertebrate genome sequences from species at least as distantly related as chicken, with greater than 80% nucleotide identity within some intervals. We sought to determine whether conservation in lncRNA sequence also extends to conservation in the expression of these lncRNAs among diverse vertebrate species. In order to identify orthologs in other vertebrates, we aligned genomic sequences orthologous to each lncRNA locus from species ranging from frog to human, and including birds and marsupials (see Materials and methods; Figures 2b, 3b and 4b).
Each lncRNA locus and its closest flanking protein-coding genes show conserved synteny across amniotic species from mouse to chicken, and a portion of each mouse lncRNA locus aligns to all the genomic sequences we analyzed (Figures 2a, 3a and 4a). The patterns of nucleotide conservation for these lncRNA loci exemplify the more general trends we observed for all such loci, including greater conservation near exon boundaries (Figure 1a). In these respects, these lncRNA loci differ markedly from protein-coding genes, which typically contain more uniformly distributed and strong conservation within exons [31].
AK043754
Blocks of aligned sequence with at least 70% nucleotide identity across all the examined amniote species are restricted to the 3' end (approximately 500 bp) of AK043754 (Figure 2). We could find no evidence of AK043754-aligning sequence within non-amniote vertebrate genomes, suggesting that this locus has either evolved extremely rapidly or originated within the amniote lineage after divergence from other vertebrates. The sequence of the putative proximal promoter, presumed to reside within the 400 bp upstream of the TSS, aligns to orthologous sequences in metatheria and eutheria; such orthologous sequence could not be identified in monotremata (platypus) and non-mammalian vertebrates. Finally, a polyadenylation signal (ATAAA) located 30 bp upstream of the 3' end of AK043754 in mouse is present in all examined amniote sequences.
Guided by the multi-species sequence alignments, we cloned the AK043754 orthologs from opossum and chicken poly(A)-selected reverse-transcribed cDNA. As illustrated in Figure 2c, the orthologous opossum and chicken sequences (as well as the orthologous zebra finch sequence [GenBank: DQ213170]) align to the mouse AK043754 sequence. Based on BlastN local alignments, the opossum (1,307 bp), chicken (1,912 bp), and zebra finch (938 bp) transcripts share approximately 38%, 29%, and 29% nucleotide sequence identity with the mouse transcript, respectively. Consistent with the multi-species genome sequence alignment, each transcript has a unique (non-aligning) TSS (indicated by grey arrows), but harbors a conserved poly(A) signal (red band) and 3' end. As with mouse AK043754, the examined orthologs lack long or conserved ORFs, indicating that this locus is unlikely to have possessed protein-coding capacity over the span of amniote evolution.
AK082072
Orthologous sequences in each of the 16 vertebrate genomes we examined (with one exception - see below) aligned to the proximal promoter and first exon of mouse AK082072 with sequence identities exceeding 85% (Figure 3b). Notably, a 5' consensus splice-site sequence (MAG|GTRAG) for U2 introns in pre-mRNA is constrained. However, sequence conservation of the second exon, including an adjacent 3' AG acceptor site and poly(A) signal, is detectable only in mammals, suggesting that this region might have arisen within the mammalian lineage after divergence from other amniotes.
AK082072 orthologs were identified in frog (754 bp), chicken (759 bp), and human (553 bp) ([GenBank: CX847574.1, CR35248.1, DA317999.1], respectively) from a BLASTn query of the NCBI (nr/nt) database. In addition, we cloned and sequenced the full-length (725 bp) opossum ortholog from poly(A)-selected reverse-transcribed cDNA. Based on the resulting BLASTn alignments, we found that the frog, chicken, opossum, and human sequences share approximately 11%, 21%, 53%, and 67% sequence identity, respectively, with their mouse ortholog (Figure 3c). Consistent with the multi-species genome sequence alignment, all transcripts utilize a conserved 5' donor site. By contrast, only the mammalian transcripts use the predicted 3' acceptor site and terminate immediately after the predicted poly(A) signal (depicted as blue and red bands, respectively, in Figure 3c).
While the relative structure of the first and last exons is conserved across therian mammals, the opossum and human orthologs contain an additional and non-homologous central exon, in each case buttressed by non-conserved AG/GT acceptor/donor sites and residing within poorly constrained genomic sequence. In fact, the opossum middle exon lies within a genomic region containing a MAR1 element (a tRNA-derived SINE (short interspersed element) specific to M. domestica [45]).
The terminal mammalian AK082072 exons lack demonstrable homology with those in the chicken and frog orthologs (Figure 3b). The second exon in chicken AK082072 is transcribed from an evolutionarily conserved region that shares >70% sequence identity with the orthologous mouse sequence (highlighted in grey) across 200 bp and harbors a poly(A) signal with 100% sequence conservation in all examined vertebrates except zebra finch. While suggestive of a highly conserved exon, we were unable to clone similar splice variants from either mouse or opossum cDNA. In contrast, the second exon of frog AK082072 appears to be specific to amphibians and, like opossum AK082072, includes a repeat element, in this case a X. tropicalis DNA transposon hAT.
AK082467/Rmst
AK082467 and Rmst orthologs from human to frog also exhibit >70% sequence identity over their proximal promoters, first exons, and 5' splice donor sites (Figure 4b). In all examined eutherians, we identified putative two-exon AK082467 orthologs that share a TSS, splice site, and exonic structure. While genomic regions containing the second exon of AK082467 share at least 60% sequence identity among the examined vertebrates, the non-eutherian vertebrates lack an upstream 3' acceptor site; hence, we expected either unspliced or differentially spliced orthologs in these species. Indeed, we cloned unspliced and differentially spliced AK082467 orthologs from chicken (30% sequence identity) and opossum (26% sequence identity) cDNA, respectively, each sharing similar 5' and 3' ends with mouse AK082467 (Figure 4c). The opossum AK082467 3' acceptor site is not conserved, as it aligns approximately 10 bp upstream of that in mouse, although this may reflect inaccuracies in the sequence alignment. Chicken AK082467 contains an additional approximately 200-bp stretch that spans the mouse intronic region. Importantly, the identified mammalian intron in AK082467 (approximately 320 bp), which is almost entirely composed of simple repeats, is not alignable to chicken or to other non-mammalian vertebrate genomes. Also, we were unable to identify a poly(A) signal within the AK082467 orthologs despite the fact that the transcripts were derived from poly(A)-selected cDNA, suggesting that the isolated transcripts were either unpolyadenylated contaminants within our cDNA samples or that the transcripts are recapped derivatives of larger RNA molecules.
Our multi-species sequence alignment (Figure 4b) revealed that only exons 1, 4, and 11 of mouse Rmst share the same exonic structure (including alignable donor and acceptor splice sites) across the examined vertebrates. At least one >50-bp stretch of >60% sequence identity resides within each of these exons. Sequences of the remaining mouse exons align to regions of varying sequence conservation among mammals, suggesting relaxed evolutionary constraint on their structures. Accordingly, we predicted vertebrate Rmst orthologs containing at least three conserved exons and a variable number of total exons. Of note, we also identified a eutherian-specific poly(A) signal residing approximately 25 bp upstream of the termination site within the mouse transcript, suggesting that other eutherians also share the same transcription stop site.
We cloned and sequenced the chicken and opossum Rmst orthologs, which contain four and seven exons, respectively. While we only identified one splice variant for each species, alternative transcripts could exist. Alignment of the identified orthologs along with the mouse and human [GenBank: NR_024037] Rmst sequences revealed striking conservation of the structures of exons 1, 4, and 11 and of the sequences of exons 1 and 11 (Figure 4c). In contrast, the mouse, opossum, and chicken Rmst exon 4 orthologs share <50% sequence identity. Furthermore, the overall sequence identity, calculated by BLASTn, between mouse Rmst and the chicken, opossum, and human orthologs is only 4%, 7%, and 22%, respectively.
Expression of selected lncRNA orthologs in the developing brain
Given the evidence that lncRNA orthologs are transcribed in diverse species, we next sought to determine whether the tissue pattern of transcription is similarly conserved. Indeed, we identified numerous homologous ESTs and cDNAs from nervous system tissue isolated from diverse species (human to zebra finch; Table 1).
Table 1. AK043754, AK082072, and AK082467 orthologs among vertebrates
To observe lncRNA expression at a finer resolution, we performed in situ hybridization of mouse, opossum, and chicken brains harvested at early and late embryonic stages, using probes specific to approximately 300-bp portions of phastCons conserved elements within AK043754, AK082072, and AK082467 exons. While the expression patterns of the lncRNA orthologs are not identical among these species, we encountered evidence of spatio-temporal regulation for each locus, with transcription typically regionally restricted within embryonic and neonatal brain tissue. Many of these regions have been implicated in the evolution of the mammalian cerebral cortex [46,47].
Probes specific to chicken, opossum, and mouse AK043754 orthologs hybridize to the germinal zone of the telencephalic cortex in coronal and sagittal sections of early developmental brain in all three species (red arrowheads in Figure 6a). While the neuroanatomical homology relationships between mammalian and avian brains remain controversial (see [46] for a review), most researchers agree that the telencephalic germinal zone is a source of neural progenitors in both mammals and birds [48]. We found that AK043754-expressing cells appear to migrate radially away from the ventricular germinal zone to the pial surface as development progresses in all three species. At later developmental stages (E12, P20, and P0 in chicken, opossum, and mouse, respectively), AK043754 is expressed within the piriform (olfactory) cortex (black arrowheads in Figure 6a). This conserved expression pattern - from the telencephalic germinal zone to a specific cortical substructure - implies negative selection acting on as yet unidentified AK043754 regulatory elements.
Figure 6. Conservation of lncRNA expression in developing avian and mammalian brains. (a-c) Digoxigenin-labeled riboprobes complementary to lncRNAs AK043754 (a), AK082072 (b), and AK082467 (c) were hybridized to chicken (E4, 6, 8,12), opossum (P12, 20), and mouse (E13, 15, 17, 18 and P0) brain sections. (a) AK043754: strong hybridization seen in the germinal zone of the telencephalic cortex at early developmental time points (red arrowheads) and then concentrated within the piriform (olfactory) cortex at later stages (black arrowheads). (b) AK082072: hybridization signals seen in the stria terminalis (red arrowheads) and the telencephalic ventricular zone (green arrowheads). Signal was undetectable at later developmental stages. (c) AK082467: hybridization signals seen in the ventricular zone of the hippocampal formation (green arrowheads), the preoptic area of the hypothalamus (red arrowheads), and the epithalamus (black arrowheads). Signal was undetectable at later developmental stages. Scale bars = 200 μm.
Early in development, chicken, opossum, and mouse prominently express AK082072 within the stria terminalis, a fiber bundle connecting the amygdala to the hypothalamus and other basal telencephalic regions, and the telencephalic ventricular zone (red and green arrowheads in Figure 6b). This expression is reduced at later developmental stages in all three species, suggesting that the locus has retained temporal in addition to spatial regulatory elements during amniote evolution.
The clearest example of a conserved expression pattern among chicken, opossum, and mouse is seen for AK082467, which hybridizes specifically to the ventricular zone of the hippocampal formation (green arrowheads in sagittal brain sections in Figure 6c), an area rich in Wnt signaling among vertebrates [49]. We also found modest conservation in expression within the preoptic area of the hypothalamus among birds and mammals and within the thalamus among mammals.
Discussion
The application of new DNA sequencing technologies over the past decade has revealed that the vertebrate transcriptome is extensive, complex, and developmentally dynamic [5]. Most components of this interleaved network of transcripts appear to have little protein-coding capacity, and their general contribution to phenotype has often been questioned. In light of the evolving definition of a 'gene' [50,51], we argue that the lncRNA transcriptional products we characterized here exhibit signatures of evolutionary constraint on sequence and transcriptional regulation that are similar to, although less pronounced than, those for protein-coding genes. These lncRNA loci thus are biologically relevant, and should be considered genes.
Conservation of lncRNA sequence
Reinforcing previous observations [6,16,18], our analyses of vertebrate phastCons scores across lncRNA transcriptional units revealed substantial evidence for more stringent purifying selection within proximal promoter sequences than within the transcripts themselves. Exemplifying this trend, the inferred promoter regions of AK082072 and AK082467 are highly conserved across vertebrates, with only punctuated conservation across the primary transcript sequences. Nevertheless, and in contrast to coding sequence, exonic conservation was observed to be <30% and was as low as 4% (for Rmst) between confirmed chicken and mouse orthologs.
Multi-exonic lncRNA loci were found to exhibit greater evolutionary constraint within exons than within introns (Figure 1a). This observation is consistent with the functionality of RNA molecules transcribed from such loci rather than, for example, functionality being imparted by the act of transcriptional elongation and chromatin remodeling. It is notable that constraint tends to be lowest on bases furthest from exon boundaries (Figure 1a). This tendency has previously been noted for protein-coding exons, where it has been associated with reduced rates of nucleotide substitution within intron-proximal exonic splicing enhancers [52]. However, lower constraint within the central portions of exons may also reflect the insertion of large transposable element sequences, which are generally free of selective constraint [53] within lncRNA exons in early eutherian evolution. In this model, large insertions into exons result in functional sequence becoming closer (in terms of fractional exonic size) to intron-exon boundaries.
Mammalian and bird AK082072, Rmst, and AK082467 orthologs share some, but not all, splice sites, exons, and introns (Figures 3c and 4c). Multi-species genomic sequence alignments of these loci revealed 100% sequence conservation across all examined vertebrates within a subset of donor and acceptor splice sites. Consensus splice-site motifs adjacent to exon boundaries were found to be under particularly strong constraint, as we found previously [18]. This indicates that rather than the opportunistic use of incidental splice sites by the splicing machinery, the presence and location of splice sites are evolutionarily conserved and likely to be relevant to the function(s) of these lncRNA loci.
Conservation of splice-site location may also demarcate an intron containing functional modules with secondary structures (such as primary miRNAs (pri-miRNAs)). As previously reported [17], lncRNA loci are enriched in Evofold-predicted RNA secondary structures. Two miRNAs (eutherian-conserved MIR1251 and vertebrate-conserved MIR135A2) are embedded in introns of Rmst alternative splice variants, indicating that this lncRNA might function as a miRNA host transcript. Similarly, numerous Evofold-predicted RNA secondary structures, which could represent as yet undiscovered miRNAs, lie within the single AK082072 intron.
Conservation of lncRNA transcription
The identification of transcribed AK082072, Rmst, AK082467, and AK043754 orthologs in birds and mammals provides strong evidence for their functionality over the 310 million years since these lineages last shared a common ancestor. Over this time span, however, it appears likely that considerable evolution of each lncRNA locus has occurred. TSSs, exon structures, and poly-adenylation signals are not always well-conserved (Figures 2c, 3c, and 4c). The structure of the AK043754 locus, for example, appears to have been altered considerably because its proximal promoter sequence in mouse is not conserved with that in chicken (Figure 2b).
We also observed similar spatio-temporal expression patterns of each lncRNA locus among distantly related vertebrates. Far from being the result of spurious transcription, the expression of these lncRNAs might instead be tightly regulated by conserved transcription factors. Indeed, Rmst transcript levels are significantly reduced in Pax2-deficient tissues [44] and AK043754 has recently been reported as a direct target of the homeobox transcription factor Nanog, which is critical for embryonic stem cell pluripotency [54]. Furthermore, a described mid-hindbrain enhancer element [55] lies within an intron of AK082072 (Figure 3b), although whether this element facilitates expression of AK082072 or a neighboring protein-coding gene remains unknown.
lncRNA functions
The observed conservation in the sequence, transcription, and expression of these lncRNA loci over hundreds of millions of years of evolution indicates that these genes must confer important functions across diverse vertebrates. Because the transcription of each of these lncRNAs is largely limited to the developing nervous system in distantly related vertebrates (Table 1), the transcripts could play critical roles in neurogenesis and neuronal differentiation in specific sectors of the developing telencephalon. The underlying molecular mechanisms could, as discussed above, involve the generation of precursor short RNAs, including pri-miRNAs. Sequence-conserved and brain-expressed lncRNA loci tend to be located adjacent to protein-coding genes that are also brain-expressed and are involved in transcriptional regulation or in nervous system development [17]. Many such lncRNA loci may thus be involved in the cis-regulation of neighboring protein-coding transcription factor genes [17,21]. Consequently, establishing whether expression of AK082072 transcriptionally regulates Mef2C (Figure 3a), a gene implicated in autism and intellectual disability phenotypes [56,57], warrants detailed investigation.
The study of lncRNAs in cortical development and evolution reflects relatively uncharted territory. Several transcription factors are expressed at specific times and regions during telencephalic development and cerebral cortex formation [58,59]. We hypothesize that slight differences in vertebrate developmental programs established during evolution are responsible for the radial expansion, which contributed to increased lamination of the mammalian cortex and, later, to the tangential expansion of cortical surface area that ultimately produced the human cerebral cortex [46,60,61]. The differential expression of lncRNA genes in a specific spatiotemporal pattern may promote neuronal diversity [62]. It is an exciting challenge to determine whether the lncRNAs evolved to differentially modulate the expression of relevant transcription factors or to act independently during telencephalic development and evolution. Our study represents an important first step by demonstrating that lncRNAs are conserved with respect to transcription, exon structure, and brain tissue-specific developmental expression during embryonic and early postnatal stages.
Conclusions
Initially selected for their extensive overlap with phastCons-predicted conserved elements and mouse brain-specific expression, the three murine lncRNA loci we examined in this study exhibit several indicators of transcript functionality. Despite a lack of extensive primary sequence conservation across amniotes, we successfully identified AK043754, AK082072, AK082467, and Rmst lncRNA orthologs with modest evolutionary constraint of exon-structure and spatio-temporal transcriptional regulation in distantly related amniotes spanning at least 310 million years of evolutionary divergence. The regulatory control of transcription and splicing patterns, evolutionary conservation of exon structure, stability of mature transcripts, and presence of predicted secondary structures suggest that the transcriptional products from each locus are functional, and should therefore be considered genes. Furthermore, similarities of spatiotemporal expression patterns for these transcripts in therian and avian developing nervous systems suggest that these lncRNA loci might contribute to neurogenesis and/or neuronal differentiation programs. Experimental inquiry of these lncRNAs will hopefully elucidate their roles in vertebrate brain development and evolution.
Materials and methods
Multi-species sequence alignments
Regions orthologous to AK043754, AK082467, Rmst, and AK082072 (including 100 kb on either side) of the following whole-genome assemblies [63] were used in this study: frog (Xenopus tropicalis; xenTro2), chicken (Gallus gallus; galGal3), songbird (Taeniopygia guttata; taeGut1), lizard (Anolis carolinensis; anoCar1), platypus (Ornithorhyncus anatinus; ornAna1), opossum (Monodelphis domestica; monDom4), mouse (Mus musculus; Mm9) rat (Rattus norvegicus; Rn4), guinea pig (Cavia porcellus; cavPor3), marmoset (Callithrix jacchus; calJac1), macaque (Macaca mulatta; rheMac2), orang utan (Pongo abelli; ponAbe2), human (Homo sapiens; Hg18), chimpanzee (Pan troglodytes; panTro2), horse (Equus caballus; equCab1), dog (Canis familiaris; canFam2), and cattle (Bos taurus; bosTau3) (Figures 2, 3 and 4; coordinates provided in Table S1 in Additional file 2). We additionally used deep sequence from a chicken BAC [GenBank: AC192716] to fill a gap in the chicken whole-genome assembly. The liftOver program [64] was used to identify orthologous regions in all non-mouse species listed. We used TBA (Threaded Blockset Aligner) to generate multisequence alignments as described previously [65], and then visualized each alignment with the program Gmaj (Generalized Multiple Alignments with Java) [66]. We used evolutionarily conserved regions (ECRs; defined as genomic segments at least 100 bp in size with at least 70% sequence identity between mouse and chicken) within and between the flanking protein-coding genes as anchors to facilitate the generation of multi-species sequence alignments [67]. Finally, percent sequence identity plots across all species considered in each alignment were graphed with the program SinicView (Sequence-aligning INnovative and Interactive Comparison VIEWer) [68].
Additional file 2. Table S1: genome coordinates used in multi-species sequence alignments. Table S2: PCR primers used for amplification of in situ hybridization probes and 3' and 5' lncRNA ortholog RACE.
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cDNA preparation, RACE and sequencing of lncRNA orthologs
Total RNA was extracted from whole brains removed from mouse (E17), chicken (E8), and opossum (P12) using RNAeasy miniprep kit (Qiagen, Hilden, Germany) and then treated with DNAse (Roche, Basel, Switzerland). Poly-A selected RACE-ready first-strand cDNA was then generated from each RNA sample (1 μg) with the GeneRacer kit, according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA). To obtain full-length 5' and 3' ends of opossum and chicken lncRNA orthologs, RLM-RACE (RNA ligase-mediated rapid amplification of cDNA ends) was performed with the opossum or chicken cDNA as template, and GeneRacer (Invitrogen) and gene-specific primers designed near the predicted 5' and 3' ortholog ends. Nested PCR of the RACE products was performed if needed. The resulting RACE products were cloned into the PCR4-TOPO vector (Invitrogen) and the inserts were sequenced. Using sequence information obtained from 5' and 3' RACE, PCR amplification and sequencing were performed with primers spanning the remaining portion of each ortholog. All primer sequences can be found in Table S2 in Additional file 2. Finally, the overlapping sequence fragments were merged into the predicted full-length cDNA with the program SeqMan (DNAStar, Madison, WI, USA). Identified lncRNA ortholog cDNA sequences were deposited into GenBank as follows: AK043754 chicken ortholog [GenBank:GU951674], AK043754 opossum ortholog [GenBank:GU951677], AK082072 opossum ortholog [GenBank:GU951678], AK082467 chicken ortholog [GenBank:GU951675], AK082467 opossum ortholog [GenBank:GU951679], Rmst chicken ortholog [GenBank:GU951676], and Rmst opossum ortholog [GenBank:GU951680].
Tissue preparation
All animal procedures were approved by the local Ethical Review Committee and performed under license from the UK Home Office (Scientific Procedures Act, 1986). Embryonic (E11, E13, E15, and E17) and postnatal (P0, P3, and adult) mice (M. musculus); embryonic (E4, E6, E8, and E12) chicken (G. gallus), and postnatal (P4, P12, and P20) opossum (M. domestica) were also used. Mouse embryos were obtained by caesarean section of time-mated pregnant dams sacrificed by cervical dislocation. Chicken embryos were anesthetized on ice and then extracted from their shells. Postnatal animals were anesthetized either on ice or by pentobarbital intraperitoneal injection (45 mg/kg). Following anesthesia, animals were decapitated, and the heads or brains were immediately embedded in Tissue-Tek embedding compound (Ted Pella, Redding, CA, USA), frozen on dry ice, and then stored at -80°C. For in situ hybridization studies, frozen sections (10 to 15 mm) were cut with a cryostat (Leica, Wetzlar, Germany) and mounted onto Superfrost Plus slides (Thermo Fisher Scientific Inc., Waltham, MA, USA).
In situ hybridization
For generation of in situ hybridization probes, universal degenerate oligonucleotide primers were designed from the most evolutionarily conserved regions of the selected mouse lncRNA loci and then PCR was performed using chicken, opossum, or mouse cDNA as template (primer sequences listed in Table S2 in Additional file 2). PCR products were cloned into the PCR4-TOPO vector (Invitrogen) and then sequenced to confirm authenticity. Sense and antisense probes were generated from selected PCR4-TOPO clones using T7 and T3 RNA polymerases and labeled with digoxigenin (DIG; Roche). Tissue frozen sections were postfixed with 4% paraformaldehyde in phosphate-buffered saline, deproteinized with 0.1N HCl for 5 minutes, acetylated with acetic anhydride (0.25% in 0.1 M triethanolmine hydrochloride), and prehybridized at room temperature for at least 1 hour in a solution containing 50% formamide, 10 mM Tris (pH 7.6), 200 μg/ml Escherichia coli tRNA, 1× Denhardt's solution, 10% dextran sulfate, 600 mM NaCl, 0.25% SDS, and 1 mM EDTA. Sections were then hybridized in the same buffer containing the DIG-labeled probe overnight at 65°C. After hybridization, sections were washed to a final stringency of 30 mM NaCl/3 mM sodium citrate at 65°C and detected using anti-DIG-alkaline phosphatase (Roche), essentially as described previously [69]. Sense probe hybridizations (Additional File 1) were used as background controls when analyzing corresponding antisense probe hybridizations.
Abbreviations
BP: base pair; DIG: digoxigenin; E: embryonic day; ECR: evolutionarily conserved region; EST: expressed sequence tag; LNCRNA: long noncoding RNA; MIRNA: microRNA; NCRNA: noncoding RNA; ORF: open reading frame; P: postnatal day; PRI-MIRNA: primary microRNA; RACE: rapid amplification of cDNA ends; RMST: rhabdomyosarcoma 2 associated transcript; TBA: Threaded Blockset Aligner; TSS: transcription start site.
Authors' contributions
RAC and LG performed the bioinformatic analyses and multi-species sequence alignments; RAC, TS, and PLO contributed to the in situ hybridizations; RAC carried out the RACE experiments and prepared the manuscript with assistance from KED, EDG, ZM, and CPP. ZM, CPP, EDG and RAC designed and coordinated the study. All authors read and approved the final manuscript.
Acknowledgements
Leah Krubitzer and Sarah Karlen (UC Davies), and Helen Stolp, Carl Joakim Ek and Norman Saunders (University of Melbourne) for M. domestica tissue; Jo Begbie (University of Oxford) for G. gallus tissue; Lisa Bluy (University of Oxford) for histological assistance; Juan Montiel (Pontificia Universidad Católica de Chile) for comments on G. gallus expression patterns, Darryl Leja and Julia Fekecs (NHGRI) for assistance with figures; Shih-Queen Lee-Lin (NHGRI) for technical assistance; and Shurjo Kumar Sen and Belen Hurle (NHGRI) for critical reading of the manuscript. RAC was supported by an NIH-Oxford Graduate Studentship in the laboratories of EDG and ZM. The project was supported from a BBSRC Project Grant BB/F003285/1 to ZM in collaboration with EDG, KED and CPP, and a BBSRC Research Grant BB/F007590/1 to CPP. This work was also supported in part by the Intramural Research Program of the National Human Genome Research Institute of the National Institutes of Health, the UK Medical Research Council, and the European Research Council (DARCGENs).
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Changes between Version 1 and Version 2 of Ticket #6561, comment 3
Ignore:
Timestamp:
Feb 12, 2012 5:59:38 PM (15 months ago)
Author:
skyper
Comment:
Legend:
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• Ticket #6561, comment 3
v1 v2
1 Found another warning where line wrapping is needed.
1Found another warning where line wrapping is needed.(r4918)
22
33If you merge nodes and because of that deleting a way which is a member of a relation you get following message:
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[Date Prev][Date Next][Thread Prev][Thread Next][Date Index][Thread Index]
[libreoffice-design] Let's work on color handling
Hi everyone,
On today's IRC chat, we approved the idea workflow [1].
This week is going to be the call for proposals for the color handling wiki
[2], so please submit or refine your proposals. The deadline for proposals
is next week's IRC chat, again at 16:00 GMT on Saturday.
[1] https://wiki.documentfoundation.org/Design/Whiteboards/IdeaWorkflow
[2] https://wiki.documentfoundation.org/Design/Whiteboards/Color_Handling
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Re: [libreoffice-design] Let's work on color handlingCor Nouws <oolst@nouenoff.nl>
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BME103:T930 Group 11 l2
From OpenWetWare
Revision as of 03:22, 29 November 2012 by Tony Nguyen (Talk | contribs)
Jump to: navigation, search
BME 103 Fall 2012 Home
People
Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
Course Logistics For Instructors
Photos
Wiki Editing Help
Contents
OUR TEAM: Group 11
Name: Benjamin Hook
Protocol
Name: Jacqueline Janssen
Protocol
Name: Samantha Boccasini
Open PCR machine engineer
Name: Tony Nguyen
R&D
Name: Ryelle Pattuinan
R&D
Name: Kenze Caulfield
Open PCR machine engineer
LAB 2 WRITE-UP
Thermal Cycler Engineering
System Design
Our re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski. Our new design incorporates some new designs such as software, screen zize, number of testing tube lots, as well as size of heating lid. All of these alterations are made to make the Open PCR system more efficient in terms of its operating system and user-friendly features.
Key Features
Our most major change to the Open PCR System is the change we made to the read-out screen on the top of the device near the heating lid. This change actually affects a few major components of our system. Not only did we move the screen to the side of the machine, rather than the top, but we also optimized the size of it. This size-change allows users to see the read-outs clearer. We also eliminated the need for a computer (or any outside device, that is) as this new larger screen will also be able to control the machine. Now the user is able to input cycles, temperature, etc. right on the screen instead of needing to plug it into a separate system. This allows for better portability and easier use.
We also changed the space of the testing tubes so now more tubes can be tested at once. To do this we lengthened the plate as well as the heating lid entirely across the top of the machine. Removing the screen from this part of the Open PCR System also allowed for this change.
Instructions
The same type of instructions to setting up the PCR is the same however running the cycle will be quite different. There will no longer be a USB drive or cord to connect it the laptop. So now the actual readings and programs will all be on the touch-screen. So now you will be able to set up the machine very similarly as before except everything will be on the screen. The idea for the new PCR machine is to be more mobile and accessible to users. Setting up the cycling will all be the same with the same options as well, so there will not be any huge differences other than it will be less worry about connection to the laptop.
Protocols
Materials
Supplied in Kit
Open PCR Machine
(32) Plastic Test Tubes to Fit into PCR Machine
PCR Power Adapter
USB Cable
Flourimeter
(8) Glass Slides
SYBR Green (200 mL)
Measurement Buffer (800 mL)
Black Box
Positive Sample (50 mL)
Eppendorf Tubes (40)
Supplied by User
Low Retention Adjustable Pipette with Disposable Pipette Tips
DNA Solution
Positive Control DNA Soltution
Negative Control DNA Solution
1 Smartphone
Pair of Gloves
1 Lab Coat
Water Sample
What the DNA Solution Should Consist Of
1 Micro-Liter Forward Primer
1 Micro-Liter Reverse Primer
50 Micro-Liter GoTaq Master Mix
.2 Micro-Liter Patient's DNA (Or Controls)
47.8 Micro-Liter Distilled Water
PCR Protocol
Step 1: Download the Open PCR Software onto Computer
Step 2: Plug in and turn on the Open PCR machine, connect the USB cable to your computer
Step 3: On the machine interface, select "DNA replication" and then choose desired number of cycles (at least 30 for quality results).
Step 4: Using the Pipette, transfer 30 samples of the patients DNA into each test tube. You should only use 1 Pipette tip for this part of the process. Also transfer the positive and negative control into separate test tubes.
Step 5: Next, transfer the forward and reverse primers into each of the 32 test tubes. 2 Pipette tips should be used in this part of the process: 1 for all of the forward primer transfers, and 1 for all of the reverse primer transfers.
Step 6: Using a new pipette tip, transfer the GoTaq Master mix into each of the 32 test tubes.
Step 7: Dilute the 32 solutions by filling the remainder of the test tube with distilled water.
Step 8: Carefully Label Each test tube with a sharpie making sure that the positive and negative controls are clearly marked.
Step 9: Open the heated lid of the Open PCR machine and place the test tubes into the designated slots. Close the lid.
Step 10: Make sure everything is properly connected and then choose "begin replication" on the interface.
Step 11: Check to make sure that the computer is correctly receiving the information, if not: stop the cycle, check the USB cord, and try again.
DNA Measurement Protocol
Step 1: When replicating is finished, remove the 32 tubes from the PCR Machine, as well as the positive and negative controls.
Step 2: Transfer DNA samples to eppendorf tube containing 800 mL of buffer. Use a new tip on the pipette for each transfer of DNA to avoid contamination. Do the same for all the controls.
Step 3: Using the rough side of the slides, place two drops over two holes on the slide of SYBR green. Carefully, using a new tip of a pipette, transfer two drops of a DNA sample on the SYBR green.
Step 4: Place the slide in the black box directly in the ray of the light.
Step 5: Take a photo of the slide using the smart phone.
Step 6: Using a new pipette tip that is marked for waste, remove the sample from the slide and dispose of it.
Step 7: Repeat steps 3-6 using a new pipette tip for each new sample or control.
Step 8: Analyze the results, knowing that the samples that glow green are the positive ones.
Research and Development
Background on Disease Markers
Primer Design
Illustration
Personal tools
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Talk:CH391L/S13/BioBricksAndRegistry
From OpenWetWare
(Difference between revisions)
Jump to: navigation, search
Line 26: Line 26:
*that provided complete digestion, with few required bases outside of their recognition site.
*that provided complete digestion, with few required bases outside of their recognition site.
*that exhibited low star (nonspecific) activity
*that exhibited low star (nonspecific) activity
+
+
It might be interesting to debate the merits of these criteria both from a historical viewpoint (in 2007) versus today in the age of Gibson cloning.
Revision as of 18:36, 31 January 2013
Gabriel Wu 23:14, 27 January 2013 (EST)There's no way there's only 700 parts in the registry. If you check the history on that page you've referenced the last time it was updated was in 2006.
Gabriel Wu: Please post the video you played in class on this website. I need a place to reference 'klivers.'
Gabriel Wu 17:42, 28 January 2013 (EST): What was the first BioBrick?
Kevin Baldridge 17:49, 28 January 2013 (EST):The registry seems to be a bit outdated. It needs some serious work with user interface and curation by experienced scientists to remove duplicates, etc. Is the AddGene database similar concept with a better implementation?
Catherine I. Mortensen 16:31, 30 January 2013 (EST): Are there any known labs that have had success with the something like AddGene or the registry? It seems users have had little success according to their reviews...
• Jeffrey E. Barrick 21:06, 30 January 2013 (EST):I think there are people out there who dip into these resources occasionally to get parts or set up a new technique in lab, especially at institutions or in countries where funding is tight. Like a lot of places, fewer people leave fewer positive comments than negative ones!
Benjamin Gilman 18:36, 30 January 2013 (EST): The page says that there's no real way to measure PoPS in vivo. Where do the PoPS numbers for different BioBricks come from then?
Neil R Gottel 19:03, 30 January 2013 (EST):Biobricks should really switch to some sort of standard where if you haven't proven that a part works as advertised, then it shouldn't be on the registry. Also the current standard is behind the current technology, as noted in the "legacy" section. Does Knight et al. still stand by the biobrick standard/concept? Is there a movement to have a real, usable registry that doesn't make me want to scream after using it for five seconds? A good start would be a "premium" section, where only parts that were demonstrated to work (or at least were presented as working at the regionals and finals) would be listed.
• Jeffrey E. Barrick 20:58, 30 January 2013 (EST):One of the activities that iGEM teams can do to earn medals is to test and improve the documentation of a part from previous years. Eventually, you would hope that the most-used parts just came to the top. It would be great if you could do a "cited ref" search on a part to see how much it is being used (by iGEM teams and in scientific publications). It would provide a way to sort parts that were never used again to the bottom (and have an all-time ranking of the best parts ever). These would be interesting human-practices exercises for an iGEM team, and we would probably find that the Coliroid is the most used part ever.
Max E. Rubinson 22:13, 30 January 2013 (EST): Did anyone find out what the first part added to the registry was?
Gabriel Wu 17:33, 31 January 2013 (EST): It was asked in class why were the original BioBrick enzymes chosen. From Tom Knight's paper (section 0.8), he explicitly states that he wanted enzymes:
• that were easy to use and reliable
• worked in compatible buffer systems
• worked at compatible temperatures
• that could be heat killed
• that provided complete digestion, with few required bases outside of their recognition site.
• that exhibited low star (nonspecific) activity
It might be interesting to debate the merits of these criteria both from a historical viewpoint (in 2007) versus today in the age of Gibson cloning.
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Quotation added by staff
Why not add this quote to your bookmarks?
Governments, like clocks, go from the motion men give them, and as governments are made and moved by men, so by them they are ruined too. Wherefore governments rather depend upon men, than men upon governments. Let men be good, and the government cannot be bad; if it be ill, they will cure it. But if men be bad, let the government be never so good, they will endeavour to warp and spoil it to their turn. Penn, William
Source: WILLIAM PENN, in his Preface to the First Frame of Government [constitution] for Pennsylvania, which was formally adopted in England, April 25, 1682.The William Penn Tercentenary Committee, Remember William Penn, 2d ed., p. 81 .The committee noted that the preface was perhaps Penns best expression of his ideas of government . · This quote is about uncategorised · Search on Google Books to find all references and sources for this quotation.
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Saying what we think gives a wider range of conversation than saying what we know. Hightower, Cullen
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After serving in the U.S. Army during World War II Cullen Tower began a career as a salesman and sales trainer. He retired and moved to Florida where he began writing the quips and advice he had been compiling throughout his career. His work was published in a newspaper feature called Salty Sally and was often found in other popular magazines including Forbes and Reader's Digest. He eventually published a collection of quotations in a book entitled Cullen Hightower's Wit Kit.
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28 October 2011
27 October 2011
26 October 2011
The Africa They Never Tell You About
"Foreign aid might be defined as a transfer
of money from poor people in rich countries
to rich people in poor countries." - Douglas Casey
-click any to enlarge-
Namibia's new (presidential) 'State House'
25 October 2011
10 Timely Quotes~
'I contend that for a nation to try to tax itself into
prosperity is like a man standing in a bucket and
trying to lift himself up by the handle.'
- Winston Churchill
24 October 2011
Former Soviet Citizen Confronts Historically Ignorant OWS Boneheads re. 'Socialism'
This is classic, h/t Jammie Wearing Fool:
Dismantled Accords + Rube Goldberg =
Best Honda Commercial, Ever
This is great, if you haven't seen it...
A few years back a television commercial was produced by Honda's UK ad agency, working in concert with company engineers who painstakingly took apart two special hand-assembled Accords to create this incredible Rube Goldberg-style contraption...
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Jackson Township (Ohio)
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Jackson is a township in Stark County.
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Revision as of 03:34, 22 January 2013
See also Kansas City (Kansas)
View of Kansas City from the top of Liberty Memorial. Union Station is in the foreground, with the rest of KC's skyline in the background.
Kansas City [1] is a large, major midwestern city on the border of Missouri and Kansas. It is the largest city in Missouri with a population around 450,000 people, and more than 2 million in its metropolitan area (2005 estimate).
Contents
Understand
Kansas City is a city that tends to hide itself from tourists. Having reportedly more boulevards than Paris and more fountains than any other city in the world except for Rome, it can be a beautiful city, too. Two neighboring cities sharing the same name: Kansas City, KS and Kansas City, MO. They are known locally as KCK and KCMO. Kansas City, Missouri is the predominant city of the metropolitan area.
The suburbs are largely south of the central city, though the area north of the Missouri River (known locally as the Northland) is beginning to experience growth similar to the south. Numbering of east/west streets begins at the Missouri River with the east/west division occurring at Main Street. Westport is around 40th St., the Plaza at 47th St., Brookside at 55th, and Waldo beginning around Gregory (71st St.).
Climate
Due to the lack of any large body of water nearby, KC experiences a continental climate with large swings and extremes of temperature. Winters vary from mild to very cold, with significant snow at times, and temperatures occasionally dipping to single digits and below 0°F (-18°C). Snow accumulation occurs 3-5 times per year, on average, sometimes exceeding a foot (31 cm). KC enjoys very pleasant spring and autumn weather, but suffers hot, humid summers. It is not uncommon for the temperature to stay above 90°F (32°C) for weeks at a time, during July and August. Because of the heat, almost all buildings in KC are equipped with air conditioning. While KC has relatively high humidity, the most common weather is clear with almost completely blue skies. The majority of the rain falls in Apr-Jun, but even in these wettest months, rain is light, compared to other cities in the region.
Downtown
Downtown Kansas City is generally defined as the areas including the River Market, the central business district (the "Loop"), the Crossroads Arts District, Crown Center and Union Station.
After years of neglect and decay in the 1980s and 1990s, downtown is making a comeback. Many once-abandoned buildings in downtown have been (or are being) rebuilt into high-dollar condominiums and loft apartments. The demand for residences downtown is quite high. In October of 2007, the Sprint Center sports arena was completed, bringing a modern sports and entertainment venue to the downtown core. The Power and Light District was constructed and opened in 2008. The "P&L" District comprises 5 high rise residential and office towers, with many clubs, bars, restaurants, and shops on the lower floors. A new performing arts center south of the convention center area opened in 2011.
The "Downtown Council" [2] operates security and cleaning/maintenance crews, who keep downtown's streets clean and serve as a security presence. Downtown is one of the safest areas of the city, both day and night.
Traveling north to south, the River Market is the first neighborhood one encounters, south of the banks of the Missouri River. A burgeoning urban neighborhood, it houses many shops, restaurants, bars, and a very active farmers' market that convenes every Saturday, even in winter.
The expressways form a loop around the central business district (CBD). Crossing the north segment of the loop takes you from the River Market to the CBD. This is where Kansas City's famed skyline reaches its greatest heights, further accentuated by its position on top of a sizable hill. The CBD has the principal concentration of white-collar employment in the metro area. The world headquarters for H&R Block is located here, within the P and L District.
From here, crossing the south segment of the loop will bring a visitor into the Crossroads Arts District, which is a regional center for art, culture, and nightlife.
When crossing Pershing, Union Station and Crown Center are seen, which houses shops, restaurants, and offices, including the world headquarters of Hallmark. The nearby Liberty Memorial [3] is the world's first World War I memorial and has spectacular views from the top. Just south of Crown Center is Union Hill, another revitalized area offering some of the best skyline views in the city.
South of Downtown
A visitor enters Midtown when they cross 31st Street. A largely residential area, it is the largest single named neighborhood in the city, and actually includes a number of smaller neighborhoods, like Hyde Park, Squire Park, Sheraton Estates, Ivanhoe, Coleman Highlands, Roanoke, Volker, Westport, Southmoreland, Valentine, North Plaza, and many others.
Just south of Midtown is the Country Club Plaza, known locally as "the Plaza", an outdoor shopping center, modeled after Seville, Spain. Farther south is the Waldo/Brookside area marked by more bars and a wonderful pre-war neighborhood. Further south, the city gives way to suburban development, which continues for about 10-15 mi (16-24 km), before giving way to rural farmland and pastures.
Get in
By plane
Kansas City International Airport (IATA: MCI) [4], serves the Kansas City area and is located in the Northland. The Kansas City Area Transportation Authority [5] offers hourly service to and from the airport on the #129 bus, called the I-29 Express [6], M-F 5AM-7PM. $1.25. KCI Shuttle [7] offers a shuttle service between the airport and downtown hotels.
Charles B. Wheeler Downtown Airport (IATA: MKC) [8], serves smaller planes.
By train
Amtrak serves KC via the recently renovated and cavernous Union Station, located at Main and Pershing. KC serves as a major stop on the daily Southwest Chief [9], line which provides service between Chicago and Los Angeles. Barring delays, eastbound trains depart at 7:26AM and westbound trains depart at 10:55PM. Kansas City is also the western terminus of Amtrak's Missouri River Runner [10], which provides twice-daily service to and from St. Louis with connecting service to Chicago.
By car
Interstate 435 forms a ring around the city. Notably I-70 goes east to St. Louis and west to Denver. I-35 is a major corridor running northeast and southwest. US Highway 71 runs north and south and forms a midtown expressway, running from the I-435/I-470 interchange, in a northwesterly course toward downtown, where it joins I-29. North of the River, US 71 follows the same route as I-29. For more information about navigation in the metro area see the Kansas City Metropolitan Area Wikipedia Article [11].
By bus
• Greyhound, [12].
• KC is also serviced by other commercial coach service companies, which arrive and depart from the depot at 10th and Troost.
• Megabus, +1 877 462-6342, [13]. Low-cost bus service to KC from Chicago, St. Louis, and Columbia; fares start at $1. Buses arrive and depart from the east side of Grand Blvd. between 2nd and 3rd Sts., next to the 3rd & Grand MetroCenter.
Get around
The national map companies produce book style maps that can be purchased at many grocery and book stores. Local real estate agents and delivery drivers use a book map produced by a local company in the crossroads district but it is hard to find. The one inside the Feist directory is good and can often be obtained locally for free but the directory itself may be too bulky to carry.
Street numbers
Addresses on east-west streets are numbered from Main Street in Kansas City, MO, and on north-south streets from St. John Ave. (or the Missouri River, in the River Market area). The direction 'South' in street and address numbers is generally implied if 'N' is not specified, except for numbered 'avenues' in North Kansas City. In most of Wyandotte County, Kansas the north-south streets are numbered and the address numbers are measured from Riverview Ave.
Navigation landmarks
• The KCTV pyramid shaped tower can be seen from many parts of the city and is well lit at night. It is next to KCPT studios at the corner of 31st and Main. It is orange.
• West and North of that is the Liberty Memorial cylinder shaped tower, which overlooks Union Station.
• The twin red brick towers of American Century Investments are oriented north and south along Main at 45th St. They are just north of the Country Club Plaza. The Kemper Museum is slightly east. The Nelson Atkins Museum is east and slightly south.
• Kansas City Community Christian Church at 4601 Main, has a group of lights that shoot a beam straight up at night. Frank Lloyd Wright designed the church. It is slightly south of and across the street from the American Century Investment Towers. The Nelson Atkins is to the east and the Kemper Museum is to the north and slightly east.
• Bartle Hall has a section that looks somewhat like a north-south suspension bridge crossing over I-670 at the southwest corner of the downtown loop. It has four towers with metal sculptures on top of each tower.
• One Kansas City Place is the tallest building in KC (as well as the state). The building walls are entirely black glass and the top has a red/white/blue light ring that can be seen at night. Located on 12th and Main in downtown.
Places and notes
• Brookside refers to the Brookside residential neighborhood as well as the collection of shops between 63d and Meyer off Brookside Blvd.
• Waldo refers to the Waldo Residential District in Kansas City, Missouri near 75th St. and Wornall Rd.
• The Country Club Plaza ("the Plaza") is an upscale shopping district built by the J.C. Nichols Co. in the 1920s.
• 39th St. usually refers to the small section of West 39th St. between State Line Road and Southwest Trafficway. Often referred to as the "39th St. Corridor, " it has many restaurants, bars and shops, and is just across the state line from the University of Kansas Medical Center.
• University of Kansas Hospital (KUMED) is the corporate name of the hospital on the KU Medical Center campus.
• Benton Curve, a site of many accidents, is a curve on I-70 where it crosses Benton Ave.
• Three Trails Crossing is the new name for the former Grandview Triangle and is the intersection of three major highways: I-435, I-470, and US Highway 71 (Bruce R. Watkins Drive). In the past it was notorious for fatal accidents but improvements and upgrades on the Triangle have mostly been completed.
• Emanuel Cleaver II Blvd., named for former mayor and current Congressman Emanuel Cleaver, comprises recently renamed portions of 47th St. and Brush Creek Blvd.
• 18th and Vine Historic District contains the Negro Leagues Baseball Museum and the American Jazz Museum.
• Library District is a recently defined district around the new Central Library at 14 West 10th St.
• Strawberry Hill is a historical area in Kansas City, KS.
• Hospital Hill is the area near 23rd and Holmes. It is home to Truman Medical Centers and Children's Mercy.
• Argentine is a part of Kansas City, KS near 30th and Argentine.
• The Crossroads Arts District is a downtown neighborhood between the CBD and Union Station, centered around the intersection of 19th St. and Baltimore. It contains dozens of art galleries and is considered by many to be the center of the arts culture in the metropolitan area. Local artists sponsor exhibits there on the first Friday of each month.
• Quality Hill is an upscale residential and commercial neighborhood on top of a hill downtown, across the river from the Kansas City Downtown Airport|Charles B. Wheeler Airport.
• Washington-Wheatley is a historically African-American/black neighborhood southeast of the 18th and Vine District.
Bus service
The Metro[14] bus is feasible within the urban core, where most of the tourist destinations are located. The MAX (Metro Area eXpress) and #57 buses connect downtown, Crown Center, Westport, the Plaza, Brookside, and Waldo. There are other lines that can drop you fairly close to your door in KCMO as well as limited stops in outlying suburbs such as Kansas City, KS, Independence, Blue Springs, Lee's Summit, the Northland, etc. There service to the casinos.
Standard fare is $1.50/trip with transfers available from the bus driver that expire two hours after issue. Some lengthy and express routes may cost more. You may purchase a One Day Pass on the bus. The Day Pass is good for local service only. It is issued at the farebox and expires at midnight. Upon boarding, request a Day Pass before depositing the $3 exact change into the farebox. Most major routes use buses that are equipped with bike racks.
The JO
If you are needing bus service in Johnson Co., KS (Overland Park, Mission, Fairway, Leawood, Olathe, Lenexa, Westwood, Merriam, Shawnee) you can access "The JO" [15] bus service. It also has lines that run from downtown KCMO to Johnson County and vice versa. The stops are limited and far apart.
Service on "The JO" is limited mostly to morning rush hour, mid-day, and evening rush hour. Fares are $1.25 for trips that begin and end in Johnson Co., $1.75 for trips that go start or end in Downtown KCMO. Transfers are available. Most buses are equipped with bike racks.
See
Downtown
• Central Business District - Downtown Kansas City houses many beautiful art deco buildings as well as examples of mid-century design and modern glass towers.
• Power and Light District - New development showcasing restaurants and rowdy bars/clubs. Also see the historic Main Street Theater (now a 6-plex) which is now the Flagship theater for AMC who has their HQ in Kansas City. The theater is said to be one of the most advanced theaters in the country complete with vibrating seats.
• Historic City Market [16] - Offers shopping, dining, concerts, events and a farmers' market.
18th and Vine Historic District
• American Jazz Museum, 1616 E 18th St, [17]. Tu-Sa 9AM–6PM, Su noon–6PM. Adult $6.
• Negro Leagues Baseball Museum, 1616 E 18th St, [18]. Tu-Sa 9AM–6PM, Su noon–6PM. Adult $6.
Combination adult ticket for both museums $8, (save $4).
Union Station area
• Union Station, 30 W Pershing Rd, [19]. Has a visitor center, theatres, restaurants, shops, the Amtrak station and the following attractions. Also, the nation's second largest train station with ceiling over 100 ft (31 m) tall.
• Science City [20], Gottleib Planetarium [21] and KC Rail Experience [22]. Ticket to all 3 is $8.95.
• National World War I Museum and Liberty Memorial, 100 W 26th St, [23]. Tu–Su 10AM–5PM (4:15PM for the tower). The memorial is the world's first for WWI. Take an elevator to the observation deck at the top of the 200-ft (62 m) obelisk for spectacular views. $8 museum, $4 tower elevator, $10 both.
• Crown Center - Home of Hallmark Cards and the Hallmark Visitors Center and Museum.
Near the Plaza
• The Country Club Plaza - The nation's first shopping center designed for the automobile. It offers high-end shops and restaurants in a quaint European atmosphere. The architecture is modeled after Seville Spain and includes a replica of the Giralda Tower.
• Visitor Center, 4709 Central, +1 800-767-7700. M-Sa 10AM–6PM, Su noon-5PM.
• Nelson-Atkins Museum of Art, 4525 Oak St, [24]. Also has the Kansas City Sculpture Park. Tu–Th 10AM–4PM, F 10AM–9PM, Sa 10AM–5PM, Su noon–5PM. Art collection housed in a beautiful 1930s building and a new, critically acclaimed modern wing by Steven Holl. Free.
• Toy and Miniature Museum Just off the UMKC campus, 3-min drive from the plaza. We-Sa 10AM-4PM, Sun 1PM-4PM. $7 Adults, $5 Children 5-12.
• Kemper Museum of Contemporary Art, 4420 Warwick Blvd, [25]. Tu–Th 10AM–4PM, F Sa 10AM–9PM, Su 11AM–5PM. Free.
• Community Christian Church, 4601 Main St, [26]. Designed by Frank Lloyd Wright and visible from the Plaza.
Other
• First Fridays in the Crossroads Arts District, (gallery crawl between Downtown and Crown Center), [27]. 7PM-9PM on the first Friday of each month. This area has been called the SoHo of the Midwest by the New York Times. Many art galleries are open late on these Fridays, attracting a growing crowd of art enthusiasts. You can walk between galleries, or hop the free trolley. Summer months are typically the most busy and entertaining. Entry to most galleries is free. Some charge for refreshments.
• Swope Park Zoo
• Ward Parkway - A tree-lined Boulevard, just south of the Country Club Plaza, that takes you past stately homes. The houses are home of some of Kansas City's elite including the family that owns Russel Stover's Candies and the Applebee's family among others. The historic houses were built in many different styles, inspired by European architecture.
• Harley-Davidson Final Assembly Plant, 11401 N Congress Ave (near the airport). Free tours (1 hr) M-F 8AM-1PM. No cameras allowed.
Do
Sports
Major league sports
• Kansas City Chiefs, One Arrowhead Dr. (I-70 to exit Blue Ridge/I-435 to exit #63B), +1 816-920-9400 (fax: +1 816-924-4570), [28]. National Football League (NFL).
• Kansas City Royals, One Royal Way (I-70 to exit Blue Ridge/I-435 to exit #63B), +1 800-6ROYALS, [29]. Major League Baseball (MLB).
• Sporting Kansas City (Wiz from the team's former name of Kansas City Wizards), 1800 Village W Pkwy, Kansas City, KS (I-435 to exit #13B/#14B, continue W to 110th St. and turn left), +1 913-387-3400 or +1 888-4KCGOAL (, fax: +1 913-387-3401), [30]. Major League Soccer (MLS).
Other sports and gaming
• Ameristar Casino, 3200 N Ameristar Dr, +1 816-414-7000, [31]. 3,000 slot and video poker machines and 60 table games. Live poker room is among the largest in the area. Also has its own microbrewery in Amerisports. You can purchase their beer throughout the casino floor as well.
• Isle of Capri Casino, [32].
• Kansas City T-Bones, 1800 Village W Pkwy, Kansas City, KS (I-435 to exit #13B/#14B, continue W to 110th St. and turn left), [33]. Northern League baseball.
• Kansas City Roller Warriors, (816)809-8496, [34]. Women's flat-track roller derby. Four local teams compete March through August.
Art
• Plaza Art Fair, Country Club Plaza, [35].
• Kansas City Renaissance Festival, 633 North 130th St, Bonner Springs, KS 66012, 913-721-2110, [36].
Food
• American Royal BBQ Contest. Largest such event in the world.
• Kansas City Hot and Spicy Festival (KC Hot Spicy Fest), [37]. Cook-off's and contests. Live bands, beer available. 21+ as the night progresses.
Music
- Kansas City has a large Jazz scene, a lot of restaurants have nightly jazz players.
• The Blue Room (The Blue Room). The Blue Room has nightly musicians
• The Majestic, 931 Broadway, Kansas City, MO 64105, 816-221-1888, [38]. The Majestic has nightly musicians
• Sandstone (Sandstone). Concerts at Sandstone
• Sprint Center, 1407 Grand Boulevard, 816-949-7100, [39]. Concerts and Events at Sprint Center
• Starlight Theatre (Starlight Theatre). Concerts at Starlight Theatre
Performing arts
- Kansas City hosts the national touring companies for some of Broadway's premier shows. Some of the highlights include Wicked, Jersey Boys and Mamma Mia! [40]
• The Midland by AMC (The Midland by AMC), 1228 Main Street, Kansas City, MO 64105, 816-283-9900, [41]. Broadway Theatre at Midland Theatre
• Music Hall (Music Hall), 301 W 13th St, Kansas City, MO 64105, (816) 513-5000, [42]. Broadway Theatre at Music Hall
• Starlight Theatre (Starlight Theatre), 4600 Starlight Road, Kansas City, MO 64132, (816) 363-7827, [43]. Broadway Theatre at Starlight Theatre
Learn
• Nelson-Atkins Museum of Art, 4525 Oak Street, Kansas City, MO 64111, [44]. Wed: 10am-4pm. Thurs, Fri: 10am-9pm, Sat:10am-5pm, Sun: 12pm-5pm. Free.
• Steamboat Arabia Museum, 400 Grand Blvd., Kansas City, MO 64106, 816-471-4030, [45]. Monday-Saturday: 10am - 5:30pm, Sunday: 12pm - 5pm.
• Downtown Library.
• Truman Library, [[Independence]] (15 min drive).
Buy
• Country Club Plaza [46].
• Oak Park Mall (suburban) [47] Largest indoor mall in the region.
• Brookside Shops [48] local non-chain shops and non-chain restaurants. Grocery stores, bar, sandwiches.
• 39th Street West [49] located roughly between State Line Rd. and Southwest Trafficway.
• Halls
• Crown Center
• Great Mall of the Great Plains (in Olathe, KS)
• Zona Rosa
• 119th Street (Suburban Kansas) The area east of the Sprint HQ is home to a number of shopping complexes like Town Center Plaza. The area is also home to two new centers: Leawood Park Place and One Nineteen. Park Place is a highly dense, mixed use suburban town center catering to local retailers.
• Antiques: KC has many unique things you will not find on the coasts. The best places to poke around are estate sales and whatnot in the outlying farm areas -- they throw nothing away. Check out the River Market Antique Mall for 4 stories of vendors selling something between trash and treasure. The West Bottoms warehouses host antique and estate sale auctions on the weekends, look to the KC Star for details.
• Crossroads Arts District [50] Highly inspiring mix of shops ranging from apparel to home design. Shops are scattered throughout this gritty district of former warehouses and industrial spaces. The district is also one of the largest collections of art galleries and studios in the Midwest.
• The Legends [51].
• City Market Area: once called the River Market (and before that the River Quay back when mobsters used to end up in trunks with bullet holes). Like Times Square, they chased out the venerable old peep shows and dive bars. Now the neighborhood hosts a farmer's market, live music during the summer from major acts, and the Steamboat Arabia Museum which hosts the largest collection of prewar silver in the world -- recovered from a large paddlewheeler that went down in the 1830's.
• Method, 1529 Grand Blvd (one block south of Sprint Center), [52]. 4PM-10PM. Fine clothing for men, modern event space. From classic to modern, streetwear, prep, casual and dress, as well as high-end women's accessories. Racks of magazines and books in the lounge area, local art, and vintage furniture. $$.
Eat
Barbecue
"Who has the best barbecue in Kansas City?" is a question that causes much debate in Kansas City. Although the debate is usually in good humor, be ready for a passionate explanation which may take some time (or a light-hearted argument if asked in front of more than one person). Although the different restaurants each have their own unique flavors, they will usually have a sauce which is thicker and sweeter than offered in most other parts of the U.S.
Kansas City is also home to a barbecue dish that is rarely found outside the area, called "Burnt Ends." These are the overcooked ends and edges of a brisket, which although dry and chewy, are amazingly smoky and full of flavor (much more flavorful than any other cut). If you are feeling open-minded about your KC barbecue experience, they are definitely worth a try.
• Arthur Bryant's, 1727 Brooklyn original location, [53]. It has pictures on the wall of presidents and other famous people who have visited. The sandwich consists of a large pile of brisket and two pieces of butternut bread. One sandwich could feed three people. Ask for sauce for take-outs. Bryant's signature sauce is a vinegar based sauce with slight lower neutral overtones. There are alternate, sweeter versions of the sauce available. There is a security guard in the parking lot at night.
• B.B.'s Lawnside BBQ, 1205 E 85th St, 816-822-7427, [54]. B.B.'s Lawnside BBQ is situated in an old roadhouse in south Kansas City, and serves Kansas City-style barbecue. All the ribs, sausage and meats are slow-smoked in a 60 year old pit with apple wood. B.B.'s prides itself on having a slightly more varied list of sides than the typical establishments in the area. This often includes Cajun/Creole staples such as Boudin Balls & Jambalaya. However, BB's is best known for live Blues, Zydeco, and other various types of live music six nights a week.
• Fiorella's Jack Stack Barbecue, 4 locations, [55]. Jack Stack is considered to be the Kansas City area's "High Class" BBQ resturant (Although Jeans & T-shirts are perfectly acceptable) Jack stack offers one of the largest varieties of meats in the metro area. Choices can vary from lamb ribs, to prime rib. Jack Stack's sauce is extremely similar to Gates sauce in that it is sweet, but slightly less tangy than Gate's version. Notable sides include their hickory pit beans and cheesy corn bake.
• Gates, [56]. When you walk in, you will immediately hear: "Hi, may I help you?" So, if you're eating in, and you have absolutely no idea what to order, shout back "Beef and a half on bun", "Mixed Plate" (Mixed plate includes ribs, sliced ham & sliced turkey) or "Burnt End on Bun". Guy Fieri of the Food Network once declared the Burnt End On Bun to be his single most favorite meal in the country. Gates Sauce is world famous and widely considered to be one of the best sauces available. It is sweet & tangy with very slight vinegar undertones.
• Haywards, College Blvd and Antioch, Overland Park, [57]. Great onion rings and the BBQ sauce is one of the most unique in the city. Also known for very juicy Burnt Ends.
• LC's, 5800 Blue Pkwy (head E on Ward Pkwy from the Plaza), [58]. Though not much to look at, don't let the barred door and admittedly intimidating surroundings scare you off; specialties here include sandwiches that feed two and thick cut fries.
• Oklahoma Joe's, KC, KS, [59]. Situated in a gas station/liquor store in Kansas City KS, Oklahoma Joe's has quickly established itself as the "Pulled Pork King" of the area. Pulled pork is the self proclamed specialty, but just about all things swine are amazing. Walk around to the serving line and have some of the best ribs in KC. The Z-Man sandwich (Beef brisket and melted provolone topped with two onion rings) and the "Carolina Style" (served open-faced with cole slaw) are also favorites. Anthony Bourdain included Oklahoma Joe's in his "13 Places To Eat Before You Die" list published in GQ magazine. Lines are punishingly long during lunch, but very well worth the wait. Call in and go to the front register for a take out order.
• Rosedale, (One block W of Rainbow on Southwest Blvd).
• Winslow's, (River Market). Ask the cook what's best. The burnt ends are really good.
Diners
• Chubby's, 3756 Broadway St. Greasy spoon that attracts the after 3AM bar closing crowd.
• Town Topic, 2021 Broadway original location. 24 hr. Small diner with low prices.
• YJ's Snack Bar, 128 W 18th St. 24 hours Thurs-Sun.. 6 tables, live music in a laid-back atmosphere. Attracts a diverse crowd from the surrounding arts district.
Italian
• Anthony's.
• Cascone's.
• Cupini's.
• Garozzo's.
• Lidia's.
• V's Restaurante.
Splurge
• 801 Chophouse (801 Steak & Chop House), 71 East 14th Street (Power and Light District/Downtown Kansas City), +1-816-994-8800, [60]. 4-11. A late 1920's New York City steak house interior with leather booths, cherry wood furnishings, wooden floors, granite counter tops and high ceilings. The chef creates a fresh sheet that highlights seasonal fresh soups and salads, oysters on the half shell, five species of fresh fish, live Maine lobsters and king crab legs. The bar opens daily at 4PM with happy hour from 4-6PM on Monday - Friday. $55. (39D 05,52DN,94D 34,56DW)
• American Restaurant.
• Bluestem.
• Capital Grille, (The Plaza).
• Em Chamas Brazilian Steakhouse, 6101 NW 63rd Ter, +1 816 505-7100, [61].
• The Golden Ox, (West Bottoms near Royal Kemper Arena). Classic steakhouse.
• The Hereford House, various locations (original at 20th and Main in Crossroads Art District. (downtown location had an arson attack on 20 Oct 2008 and is temporarily closed).
• Jaspers'.
• Savoy Grill, (downtown across from Quality Hill).
• Little Chef. First export from the UK roadside Kwality Koozine Specialists.
Other
• Jess & Jim's Steakhouse, 517 E. 135th St. Kansas City, MO 64145, +1 816 941-9499, [62]. The quintessential Midwestern steakhouse. Known for the 25 oz. Playboy Strip that put this little place on the national map. USA Today declared Jess & Jim's one of the nation's top steakhouses and former US president Bill Clinton is known to stop in when he visits Kansas City.
• Stroud's. Fried chicken.
• The Peachtree, 31 E. 14th Street, (816) 886-9800, [63]. Authentic southern/soul food restaurant located in the Power and Light District. The lunch menu, at $7.99 for an entree, two sides, and bread, is a really excellent value. The restaurant serves delicious fried catfish, meatloaf, southern fried chicken, etc.
Drink
There is a popular brewery in KC by the name of Boulevard whose beers are available on tap at many different bars and restaurants throughout town.
Live Music Venues: Uptown Theatre (Midtown, on Broadway and Valentine) hosts lots of up and coming acts and many timeless performers play here because it is so famous. Beaumont Club is a soulless Westport C&W club that hosts live music but has acoustics' issues and a mechanical bull. The Grand Emporium used to be one of the best blues clubs but after being bought by the owners of the Beaumont they changed it considerably, including the name to Tao.
• John's Upper Deck, 928 Wyandotte Street Kansas City, MO 64105 (Neighborhoods: Central Business District, Greater Downtown), (816) 474-5668. (per "Rock H." on yelp.com, Four Stars) Atmosphere: When the weather is nice, the deck is the place to be. Gorgeous. Skyline, fans if needed, open air and nice conversation acoustics of course. Food: I've had a handful of items and they were all very good, except maybe the wings, but that's been a year+, so maybe they're better now. Half price appetizers most of the time. Unbeatable. Drinks. $2 Domestics. One night, before Freaker's Ball down the street at Midland, Domestic beers were $1. ONE DOLLAR! The Negatives: The deck is up what amounts to about 5 flights of stairs, and the steps were a little slick. Not a big deal normally, except the bathrooms are on the first floor. Still, not a showstopper. Karaoke. It's a super pleasant experience until the LOUD music/karaoke begins. That's when a lot of people bail out. A really solid bar and grill and unique experience. $.
Downtown
• 12 Baltimore, 12th St and Baltimore (attached to the upscale Hotel Phillips).
• Angel's Rock.
• Bice Italian Bistro.
• ChinaBar.
• Crosstown Station, (S of the Sprint Center Arena next to KC Star Bldg on McGee St). New indoor live music venue and bar. Voted Best Lights, Sound and Stage.
• The Drum Room. Historic jazz and blues location.
• Flying Saucer Draught Emporium, 101 East 13th St., [64]. Beer bar with 80 taps and many bottles. Trivia Tuesdays 7:30, Rare Beer Nights Wednesdays.
• Howl at the Moon Dueling Piano Bar.
• John's Deck.
• Kansas City Live!. Open-air live music and entertainment venue, open-container allowed. Surrounded by the Power & Light District's various bars and eateries.
• Knucklehead's, 2715 Rochester St, +1 816 483-1456, [65]. W-Su. Also known for the wide variety of music they bring to KC like country, blues, rockabilly. Roots, singer/songwriters and Zydeco. Free shuttle available.
• McFadden's Sports Saloon.
• The Peanut.
• The Phoenix. Live jazz
• The Quaff, 1010 Broadway. Large, very popular bar with scantily-clad waitresses and tough guy bartenders. Frequented mostly by the post-college crowd.
• Tengo Sed Cantina.
• Willie's. Small sports bar with a largely post-college crowd. A franchise out of Columbia, MO; a great place to go to see University of MO games.
• Zebra Room, (in Hotel Aladdin). Newly remodeled; has a swanky zebra-skin theme. Great food and a martini lounge on the mezzanine level.
Brookside
• The Brooksider.
• Carmen's Cafe, 6307 Brookside Plaza, 816-333-4048, [66]. Carmen's serves up tapas, pasta, and Latin- and Italian-style entrees, wine and cocktails in an intimate and friendly setting. Come in through the downstairs bar. $15-30.
• Charlie Hooper's.
• Sharp's 63rd Street Grill, 128 W 63rd Street, 816-333-4355, [67]. breakfast 8a-2p, lunch and dinner 11a-10p. Famous for breakfast, bar food, sandwiches, chili, and water chestnut soup. Sharp's is LGBT-friendly, but not exclusive. Bar features a rotating selection of specialty cocktails. Service is friendly and accurate, but not the speediest, so plan to relax over your meal. $10-15.
Country Club Plaza
• Blonde.
• Fred P Ott's.
• The Granfalloon.
• JJ's. JJ's is a laid back, wine and martini type bar that generally caters to older patrons, but the drink selection is good and the bartenders friendly. The outdoor patio is heated so it can be enjoyed during most of the year.
• O'Dowd's. O'Dowd's is a standard Irish Pub but features live music often and is one of the more upbeat places on the Plaza. Features an open air rooftop bar.
• Tomfooleries. Tom's (as it is frequently referred to) is a restaurant by day and bar by night. The downstairs is generally quieter and calmer than upstairs in the evening. There is also a patio area outside. The cheapest place to drink on the Plaza, as domestic beer is $2 after 9PM. A good place to start the night.
Crossroads
• Balanca's Pyro Room, 1809 Grand, 816-474-6369, [68]. 6p-3a. Balanca's prides itself on the diversity of its crowd - it's not unusual to see drag queens drinking and dancing with cowboys and hip-hop aficionados in the downstairs bar, while goths play pool in the upstairs bar with yuppies and hipsters. It's weird, but it works.
• The Brick.
• Bulldog, 17th and Main Sts.. Cocktails and fine eats served in nice atmosphere.
• The Cashew, 20th and Grand. Open air cafe feel to this two-story bar. In the summer months, the windows are raised up (think: "garage door") and both stories gain a relaxing breeze and a nice view of the city. Occasionally has live music, popular stop during First Fridays.
• Danny's Big Easy, 16th and Main Sts..
• Grinders, 18th and Locust (3blocks E of Grand St.). Eclectic beer selection and authentic Philly Cheeseteak sandwiches.
• Jilly's.
Midtown
• Davey's Uptown Rambler's Club.
• Mint Ultralounge (Formerly The Empire Room).
• The Grothaus+Pearl Gallery, [69].
• The Hangout.
• Harling's.
• The Lava Room.
• The Levee.
• The Newsroom.
• The Velvet Dog.
Northeast Industrial District
• Knucklehead's, 2715 Rochester St..
River Market
• The Cup and Saucer.
• Harry's Country Club.
• Kabal.
• Minsky's.
• Skybox.
Waldo
• 75th Street Brewery, 520 W 75th Street, 816-523-4677, [70]. 5 regular beers, a non-alcoholic root beer, and an assortment of seasonal offerings brewed on premises. The kitchen also serves up salads, sandwiches, steaks, snacks, and other tasty treats. Live music on Sunday evenings.
• Bobby Baker's Lounge.
• Fin's Waldo Bar.
• The Gaf.
• Lew's.
• Swizzle.
• Tanner's Waldo.
Westport
• America's Pub.
• Buzzard Beach.
• Dark Horse Tavern, [71].
• Dave's Stagecoach Inn.
• Karma.
• Kelly's.
• Harpo's.
• Harry's Bar and Tables.
• McCoy's Public House, 4057 Pennsylvania, 816-960-0866, [72]. 11a-3a, 11a-12a on Sundays. House-brewed beer and full menu, including vegetarian options. 6 regular beer offerings plus a rotating selection of seasonal beers. Cocktails and a selection of scotches and bourbons also available. $15-25.
• Westport Flea Market, 817 Westport Road, 816-931-1986, [73]. 11a-1a, kitchen closes at 11p. Regularly voted Kansas City's best burger, the Flea has a full food menu, but is really more about drinks and hanging out. Live music on Sundays, karaoke on Fridays and Saturdays, chess on Tuesdays, trivia on Wednesdays. Relaxed crowd, pool tables and pinball. 23 beers on tap.
• Firefly - 'A Modern Speakeasy'.
Rudy's Tenampa Taqueria (Authentic Mexican)
West Bottoms
• Korruption.
Sleep
• Homewood Suites by Hilton Kansas City-Airport, 7312 NW Polo Drive, Kansas City, Missouri, 64153, [74]. checkin: 3pm; checkout: 12pm. Features suites with fully equipped kitchens and separate sleeping areas. Complimentary perks such as full hot breakfast, Wifi and shuttle service to the airport.
• Hampton Inn Kansas City-Near Worlds of Fun, 4233 N. Corrington Avenue, 816-452-1010, [75]. Just one mile from Kansas City’s best amusement park and under half a mile from the water park, Oceans of Fun. Features free hot breakfast daily and free high-speed internet in every room.
Budget
• Microtel Inn and Suites Kansas City International Airport, 11831 NW Plaza Cir, +1 816 270-120, [76].
Mid-range
• Ameristar Casino Hotel Kansas City, 3200 N Ameristar Dr, +1 816 414-7000, [77]. Refurbished lobby, 184 rooms all with king or 2 queen size beds.
• AmeriSuites Kansas City/Airport, 7600 NW 97th Ter, +1 816 891-0871, [78]. 2 mi S of MCI Airport and 12 mi N of downtown.
• Embassy Suites Kansas City International Airport, [79]. 4 mi from MCI Airport, 1 mi from Zona Rosa.
• Southmoreland on the Plaza, 116 E 48th St, [80]. B and B.
• Courtyard Kansas City Airport, 7901 NW Tiffany Springs Parkway, +1 816 891-7500 (fax: +1 816 891-8855), [81]. 6 mi from MCI Airport. Complimentary shuttle service. (39.272635,-94.67106)
Splurge
• Courtyard Kansas City Country Club Plaza (Historic Courtyard Marriott Plaza), 4600 JC Nichols Pkwy, [82]. In the heart of the Plaza. The hotel has free wifi, complimentary parking, refrigerators and microwaves in every room and bar.
• The Hilton President Hotel, (sign reads 'President' in P and L District).
• Hotel Phillips, 106 W 12th St, [83]. Renewed through a $20 mil restoration project, a luxury boutique hotel. First opened in 1931, the hotel is on the National Registry of Historic Places, cited as a classic example of Art Deco architecture. Walnut paneling and marble floors.
• Hyatt Regency Crown Center, [84]. Newly renovated, connected to the Crown Center office and shopping complex.
• InterContinental Kansas City at The Plaza, 401 Ward Pkwy, [85]. 366 luxury-class guest rooms and suites.
• The Marriott Hotel, 12th and Wyandotte (Convention District). 22-floor hotel that connects to the historic Muehlebach Hotel. Most recognizable at night, when the front of the hotel shows a light display from dusk-11PM.
• The Raphael Hotel, 325 Ward Pkwy, [86]. An historic landmark in the Plaza, upscale boutique hotel with amenities for business, leisure and heritage travelers.
• The Westin Crown Center, 1 E Pershing Rd, +1 816-474-4400, [87]. Located within Hallmark's Crown Center, an 85-acre complex of shops, restaurants and theaters.
Contact
• Kansas City Convention & Visitor's Bureau, [88]
• Missouri Division of Tourism, [89]
Stay safe
Like most major cities, the tourist and business areas (downtown and the Plaza) are generally safe day and night, but it is always best to be aware of your surroundings. It's also best to be especially cautious during the evenings and night around the P and L District, where there has been an increase in crimes. Most violent crime happens during the late nighttime hours. Neighborhoods east and southeast of the downtown/midtown/Plaza areas are experiencing an increase in violent crime. These areas offer little for visitors and are best avoided.
KC is in Tornado Alley, so make sure you check the local weather forecasts and make sure you understand what to do during a tornado. Tornado sirens are in place to warn of incoming twisters. The system is tested every month on the first Wednesday at around noon. If you hear the sirens any other time, they're serious!
Cope
Consulates
• Germany (Honorary), 8014 State Line Ste 203, Leawood, KS, +1 913 642-5134 (, fax: +1 913 642-5348), [91].
Get out
Kansas City is one of those places where hidden in darker corners, you will find the gems, the places to visit and go. Rarely do things jump out at you and say: come visit this attraction, food, shopping, or historical establishment. Pick up a Pitch Weekly for nightlife information. If you want to relax and enjoy a walk, check out Loose Park just south of the Country Club Plaza. During the spring and fall, it's one of the most beautiful places to stroll. When you are done there are plenty of other beautiful and majestic cities in this region like Omaha, Chicago, Saint Louis, Independence, Lee's Summit, and Des Moines.
Routes through Kansas City
Council BluffsSt. Joseph N S END
Des MoinesKearney N S Kansas City, KSWichita
TopekaKansas City, KS W E IndependenceColumbia
HutchinsonOverland Park W E Lee's SummitJefferson City
AtlanticSt. Joseph N S NevadaJoplin
De SotoSt. Joseph N S Kansas City, KSTulsa
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Lawrenceville
From Wikitravel
Central Georgia : Lawrenceville
Revision as of 15:29, 20 February 2008 by 74.224.75.239 (Talk)
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Lawrenceville[1] is a city in Georgia, on the outskirts of Metro Atlanta.
History
Lawrenceville came into being on December 15, 1821 and is the county seat of Gwinnett County. Lawrenceville was named after Captain James Lawrence who was commander of the frigate Chesapeake. During a battle with H. M. S Shannon on June 1, 1813, off the coast near Boston, Massachusetts captain Lawrence was mortally wounded. His dying words were, "Don't give up the ship."
In 1978 Larry Flynt, publisher of Hustler Magazine, was shot on the Lawrenceville courthouse square allegedly by Joseph Paul Franklin.
Get in
By Plane
There is no regular passenger service into Lawrenceville but there is large general aviation airport that charter flights can fly into. Gwinnett County's Briscoe Field has a 6,000 foot runway with two FBOs supplying both 100LL and Jet-A fuel.
By Car
Travel north on Interstate 85 from Atlanta to State Highway 316. Travel east on 316 to Highway 20 and turn right.
Get around
See
• DeKalb's House before she went to midtown.
Do
Buy
Eat
• Lil' River Grill 179 East Crogan St. 770 995-1183 [2]. An American classic -,Fine dining and an extensive wine list.
• The Flying Saucer Retro Cafe & Baker], 125 W. Crogan St. 770 339-9930.[3], From soups and sandwiches to wraps, salads, quiches and desserts…coffee, espresso, specialty drinks, and more!
• Corner Stop Café, Southern Style Cookin’ in The Heart of Historic Downtown Lawrenceville, 195 N. Perry St. / (770)962-4112, Mon - Fri: 7 AM - 10 AM & 11 AM - 3 PM (Dinner Hours Fri: 5 PM - 8 PM)
Drink
Sleep
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
3235.8 - Population by Age and Sex, Australian Capital Territory, 1996
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 23/12/1997
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Irregular; ISSN:1329-4156; this publication contains final estimated resident population (ERP) by age and sex for the Statistical Local Areas (SLA) of the ACT at 30 June 1996. The ERP of an area is the estimate of the number of people who usuallyreside in that area.
This publication has been converted from older electronic formats and does not necessarily have the same appearance and functionality as later releases.
© Commonwealth of Australia 2013
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ABOUT THIS RELEASE
Provides an up-to-date overview of what has been happening in the South Australian economy and contrasts the state's performance with that of the Australian economy. Provides details of movements in a range of economic indicators and also contains time series information.
Changing to electronic format, from the Feb 2004 release.
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Research article
A genome-wide DNA methylation study in colorectal carcinoma
Muhammad G Kibriya1*, Maruf Raza2, Farzana Jasmine1, Shantanu Roy1, Rachelle Paul-Brutus1, Ronald Rahaman1, Charlotte Dodsworth1, Muhammad Rakibuz-Zaman3, Mohammed Kamal2 and Habibul Ahsan1,4,5,6
Author Affiliations
1 Department of Health Studies, The University of Chicago, Chicago, IL 60637, USA
2 Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka 1000, Bangladesh
3 Columbia University and University of Chicago Research Office in Bangladesh, Dhaka, Bangladesh
4 Department of Human Genetics, The University of Chicago, Chicago, IL 60637, USA
5 Department of Medicine, The University of Chicago, Chicago, IL 60637, USA
6 Comprehensive Cancer Center, The University of Chicago, Chicago, IL 60637, USA
For all author emails, please log on.
BMC Medical Genomics 2011, 4:50 doi:10.1186/1755-8794-4-50
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1755-8794/4/50
Received:23 March 2011
Accepted:23 June 2011
Published:23 June 2011
© 2011 Kibriya et al; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background
We performed a genome-wide scan of 27,578 CpG loci covering 14,475 genes to identify differentially methylated loci (DML) in colorectal carcinoma (CRC).
Methods
We used Illumina's Infinium methylation assay in paired DNA samples extracted from 24 fresh frozen CRC tissues and their corresponding normal colon tissues from 24 consecutive diagnosed patients at a tertiary medical center.
Results
We found a total of 627 DML in CRC covering 513 genes, of which 535 are novel DML covering 465 genes. We also validated the Illumina Infinium methylation data for top-ranking genes by non-bisulfite conversion q-PCR-based methyl profiler assay in a subset of the same samples. We also carried out integration of genome-wide copy number and expression microarray along with methylation profiling to see the functional effect of methylation. Gene Set Enrichment Analysis (GSEA) showed that among the major "gene sets" that are hypermethylated in CRC are the sets: "inhibition of adenylate cyclase activity by G-protein signaling", "Rac guanyl-nucleotide exchange factor activity", "regulation of retinoic acid receptor signaling pathway" and "estrogen receptor activity". Two-level nested cross validation showed that DML-based predictive models may offer reasonable sensitivity (around 89%), specificity (around 95%), positive predictive value (around 95%) and negative predictive value (around 89%), suggesting that these markers may have potential clinical application.
Conclusion
Our genome-wide methylation study in CRC clearly supports most of the previous findings; additionally we found a large number of novel DML in CRC tissue. If confirmed in future studies, these findings may lead to identification of genomic markers for potential clinical application.
Background
Colorectal carcinoma (CRC) is one of the most common human malignancies worldwide, and an increasing incidence of CRC in Asia has been reported [1,2]. CRC cells develop several genetic and epigenetic alterations in cancer-related genes to achieve malignant status [3]. Promoter hypermethylation coupled with loss of heterozygosity at the same locus results in loss of gene function in many tumor cells [4]. Alterations in DNA methylation in cancer, in general, have been known for 25 years, including hypomethylation of oncogenes and hypermethylation of tumor suppressor genes [5]. Identification of specific DNA methylation markers would be helpful for understanding pathogenetic mechanisms, as well as for developing new therapeutic strategies. So far most of the studies addressing DNA methylation and cancer have followed the candidate gene approach, or addressed a handful of genes, or have used cell lines [6-8]. Recently Ang et al. used Illumina's GoldenGate array covering 1,505 loci and found a total of 202 loci covering 132 genes to be differentially methylated in CRC [9]. Attempts are being made to classify CRC by methylation patterns that correlate with prognosis [10-15]. A recent study suggests that there may be a significant difference in DNA methylation profiles between cancer cell lines and original tumor tissue emphasizing the need to be cautious in using cell lines as a tumor model for molecular studies of cancer [16].
To our knowledge, there is no published study from Southeast Asia addressing these molecular features in CRC to better understand the underlying pathology. There is epidemiologic evidence of a link between ethnicity, certain food habits (more red meat, less vegetables) and CRC [17]. With very few exceptions in the tribal areas, the Bangladeshi population is relatively homogenous ethnically and has a more or less similar pattern of food habit. In this study we have used Illumina's Infinium methylation assay to study the methylation status in 27,578 CpG sites covering 14,475 genes in paired CRC and surrounding healthy tissue from Bangladeshi patients with CRC to identify differentially methylated loci involved in CRC.
Methods
Tissue Samples
The samples were collected from surgically removed colonic specimens received by the department of Pathology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh during the period of December 2009 to March 2010. All samples were collected by one surgical pathology fellow (MR) from the operating room immediately after the surgical resection. We considered the consecutive 24 eligible cases with histologically confirmed diagnosis of CRC. Histopathology was done independently by two histopathologists (MK & MR), and there was concordance in all 24 cases. For each patient, one sample was collected from the tumor mass, and another sample was taken from the resected unaffected part of the colon about 5-10 cm away from the tumor mass. Thus, from each individual we obtained a pair of tumor and normal tissues. From each site, the tissue was collected as fresh frozen and also in RNA-stabilizing buffer. The samples were shipped on dry ice to the molecular genomics lab at The University of Chicago for subsequent DNA extraction and methylation assay. Patient characteristics are shown in Additional File 1 Table S1. For each patient, we also abstracted key demographic and clinical data and tumor characteristics from hospital medical records. Written informed consent was obtained from all participants. The research protocol was approved by the "Ethical Review Committee, Bangabandhu Sheikh Mujib Medical University", Dhaka, Bangladesh (BSMMU/2010/10096) and by the "Biological Sciences Division, University of Chicago Hospital Institutional Review Board", Chicago, IL, USA (10-264-E).
Additional File 1. Table S1: Patient characteristics. Table S2: Result from Gene Set Enrichment Analysis (GSEA) and GO-ANOVA. Table S3: Differentially methylated loci (DML) in CRC compared to adjacent normal colonic mucosa. Table S4: Validation of microarray methylation data by qPCR-based methyl profiler assay of twelve genes in paired samples from 10 patients (20 samples).
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DNA extraction and quality control
DNA was extracted from fresh frozen tissue using Puregene Core kit (Qiagen, Maryland, USA). The average 260/280 ratio was 1.85. Electropherogram from Agilent BioAnalyzer with Agilent DNA 12000 chips showed the fragment size to be >10000 bp (Additional File 2 Figure S1).
Additional File 2. Figure S1: Electropherogram of DNA samples. Agilent 2100 BioAnalyzer electropherogram of 10 DNA samples (in different colors) overlaid on ladder marker (shown in violet). Size (bp) of each peak of the DNA ladder in shown on the top of each peak. The figure shows DNA fragment size >10000 bp.
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RNA extraction and quality control
RNA was extracted from RNA Later preserved colonic tissue using Ribopure tissue kit (Ambion, USA, Cat# AM1924). Quality was checked on Agilent BioAnalyzer. RNA from two patients showed poor quality and that was also reflected on the microarray data.
Genome-wide methylation assay
The Infinium Methylation-27Assay was used to detect 27,578 CpG sites genome-wide, spanning 14,495 genes. The CpG sites were located within the proximal promoter regions of genes, with the distance to transcription start site (TSS) ranging from 0 to 1499 bp averaged at 389 ± 341 bp. For bisulfite conversion, EZ DNA methylation kit (Zymo Research, USA) was used. Paired samples (CRC and corresponding normal) were processed on the same chip, and all the 48 samples were processed on 4 chips (12 samples/chip) at the same time to avoid batch effect. The Illumina protocol was followed for the methylation assay. A Tecan Evo robot was used for automated sample processing and the chips were scanned on a single BeadArray reader (S-428). Control panel in the BeadStudio analytical software showed excellent intensity for staining (above 15,000), clear clustering for the hybridization probes, good target removal intensity (<400) and satisfactory bisulfite conversion.
Validation of Infinium methylation platform by q-PCR array
We used commercially available custom Methyl profiler PCR array from Qiagen-SABiosciences, which does not require bisulfite conversion. Manufacturer's protocol (http://www.sabiosciences.com/dna_methylation_custom_PCRarray.php webcite) was followed for the assay and ABI7900 RT-PCR instrument was used to read the plates. Basically, the assay relies on differential cleavage of target sequences by two different restriction endonucleases - methyl specific (MS) and methyl dependent (MD), whose activity require either the presence or absence of methylated cytosines respectively in their recognition sequences. Details of the assay are published elsewhere [18]. Using the standard ΔΔCt method the proportion of hypermethylated and intermediately methylated DNA was calculated [18] using the manufacturer supplied Excel macro spreadsheet.
High density SNP assay
We used Illumina 610 Quad BeadChip (Illumina Inc.) to obtain the copy number data from a total of 620,901markers (592,532 SNPs and 28,369 CNV probes).
Genome-wide gene expression microarray
We used HT12 v4 BeadChip (Illumina Inc.) for gene expression. The chip contains a total of 47,231 probes covering 31,335 genes. Paired samples were processed in same chip (12 samples/chip) and all 48 samples were processed in a single batch using 4 chips to minimize batch effect.
Statistical analysis
To compare the continuous variables (e.g. number of detected loci/samples or average signal intensity/average β value etc. among the two groups), we used one-way analysis of variance (ANOVA).
Genome-wide Methylation data analysis
For measuring methylation, we used the Illumina BeadStudio software to generate the β value for each locus from the intensity of methylated and unmethylated probes. We used the intensity values with and without background normalization. The background value is derived by averaging the signals of built-in negative control bead types, which are designed to be thermodynamically equivalent to the regular probes but lack a specific target in the transcriptome. The β is calculated as (intensity of methylated probe)/(intensity of methylated probe + intensity of unmethylated probe). Hence, β ranges between 0 (least methylated) and 1 (most methylated) and is proportional to the degree of methylated state of any particular loci. The methylation module of BeadStudio was used for differential methylation analysis using Illumina custom model. The model operates under the assumption that the methylation value β is normally distributed among biological replicates corresponding to a set of biological conditions (tumor and normal in the present scenario). DiffScore of a probe is computed as:
In addition to the Illumina BeadStudio differential methylation analysis, we exported the BeadStudio generated β-values to PARTEK Genomic Suite [19] for further statistical analyses. For statistical analysis we used these β-values with and without quantile normalization. In this way, initially we examined four sets of data - (1) no normalization of signal intensity, no normalization of β-values; (2) no normalization of signal intensity to calculate β-values initially, but subsequently quantile normalization of β-values were used; (3) background normalization of signal intensity to calculate β-values, but no normalization of β-values were used; (4) background normalization of signal intensity to calculate β-values, and quantile normalization of β-values were used. Principal component analysis (PCA) and sample histograms were checked as a part of quality control analyses of the data. Mixed-model multi-way ANOVA (which allows more than one ANOVA factor to be entered in each model) was used to compare the individual CpG loci methylation data across different groups. In general, "tissue" (tumor/adjacent normal), sex (male/female) and tumor location (proximal colon/distal colon) were used as categorical variables with fixed effect since the levels "tumor/normal", "male/female", and "proximal/distal" represent all conditions of interest; whereas "case ID#" (as proxy of inter-person variation) was treated as categorical variable with random effect, since the person ID is only a random sample of all the levels of that factor. Method of moments estimation was used to obtain estimates of variance components for mixed models [20]. As per the study design, we processed both the CRC tissue and the corresponding adjacent normal sample from one individual in a single chip (one chip accommodates 12 samples) and all the four chips required to run a total of 48 samples were run in a single batch to avoid batch effect. In the ANOVA model, the β-value for the CpG loci was used as the response variable, and "tissue" (tumor or normal), case ID#, "sex" and "location" were entered as ANOVA factors. It may be noted that "sex" and "location" were nested within "case ID#". One example of a model is as follows:
where Yijklm represents the m-th observation on the i-th Tissue j-th Sex k-th Location l-th CaseID, μ is the common effect for the whole experiment, εijklm represents the random error present in the m-th observation on the i-th Tissue j-th Sex k-th Location l-th CaseID. The errors εijklm are assumed to be normally and independently distributed with mean 0 and standard deviation δ for all measurements.
In GO Enrichment analysis, we tested if the genes found to be differentially methylated fell into a Gene Ontology category more often than expected by chance. We used chi-square test to compare "number of significant genes from a given category/total number of significant genes" vs. "number of genes on chip in that category/total number of genes on the microarray chip". Negative log of the p-value for this test was used as the enrichment score. Therefore, a GO group with a high enrichment score represents a lead functional group. The enrichment scores were analyzed in a hierarchical visualization and in tabular form.
In addition to looking at differential methylation at the level of individual CpG loci, we also examined the differential methylation of "gene sets" using the Gene Set Enrichment Analysis (GSEA) [21]. Given an a priori defined set of genes S (sharing the same GO category), the goal of GSEA was to determine whether the members of S were randomly distributed throughout the ranked list or primarily found at the top or bottom. Considering the fact that GSEA can look at single variable (unadjusted β-value), we also used GO-ANOVA which offers adjustments for other factors such as "person-to-person" variation, "tissue type" variation etc.
GO-ANOVA is a mixed model ANOVA to test the methylation of a set of genes (sharing the same GO category) instead of an individual gene in different groups [19]. The analysis is performed at the gene level, but the result is expressed at the level of the GO-category by averaging the member genes' results. The equation for the model was:
where Y represents the methylation status of a GO-category, μ is the common effect or average methylation of the GO-category, T is the tissue-to-tissue (tumor/healthy) effect, P is the patient-to-patient effect, G is the gene-to-gene effect (differential methylation of genes within the GO-category independent of tissue types), S(T*P) is the sample-to-sample effect (this is a random effect, and nested in tissue and patient) and ε represents the random error.
Cross-validation
For the one-level cross validation, the data was first divided into 10 random partitions. In each iteration, 10% of samples were held out for testing while the remaining 90% samples were used to fit the parameters of the model. We also used a 6 × 10 two-level nested cross-validation [22]. In the outer cross-validation, with random 1/6-th of the samples (n = 8) were held out as test samples, and the remaining 40 samples were used in an inner 10-fold cross-validation (1/10-th of these samples (n = 4) were held out at each iteration, and it was repeated 10 times) to determine the optimal predictor variables and other classifier parameters. The model that performed the best on the inner cross-validation was applied to the 8 test samples that were held-out in the outer cross-validation. This was repeated 6 times. Thus we had a total of 10 × 6 or 60 permutations for inner cross-validation and 6 for the outer cross-validation. The inner cross-validation was performed in order to select predictor variables and optimal model parameters, and the outer cross-validation was used to produce overall accuracy estimates for the classifier. Initially we tested several classification methods: (a) K-Nearest Neighbor (KNN) with Euclidean distance measure and 1-neighbor, (b) K-Nearest Neighbor (KNN) with Euclidean distance measure and 3-neighbor (c) nearest centroid with equal prior probability and (d) linear discriminent analysis with equal prior probability. Based on the results (normalized correct rate), we finally used KNN with Euclidean distance measure and 3-neighbor as the classifier, and regarding the number of variables, we tested 1 through 50 variables. For automated variable selection, we used 2-way ANOVA where tissue type and case ID# were used as ANOVA factors. One of the loci with maximum delta-β (DAB2IP) was forced into the model.
Genome-wide Copy number (CN) analysis
BeadStudio normalized intensity values were imported into PARTEK genomic suit [19]. Intensity data from the normal tissue was used as reference for generating copy number data for each marker. Standard PCA and sample histogram were generated as part of QC. Genomic segmentation was done with a setting of minimum of 10 markers, p-value threshold of 0.001 for two neighboring regions having significantly differing means. A segment was considered as amplification if the mean CN was ≥2.5 and deletion if the mean was ≤1.5. The phenotype (CRC or normal) was tested for association with amplification/deletion status of the sample using Pearson's Chi-square test.
Genome-wide Gene Expression analysis
In BeadStudio, quantile normalization was used for the intensity data. PCA detected the arrays from the same two patients as outlier which showed poor RNA quality on Agilent BioAnalyzer. Microarray data from those two patients were excluded from the analysis. Differential gene expression analysis was done using the same mixed model multi-way ANOVA [19] as in case of methylation analysis. We report genes to be differentially expressed only if that shows at least 1.3 fold change in either direction at FDR 0.05. We used this 1.3 fold as cut-off based on the power calculation from our data. Given the sample size, we had 80% power to detect 90% of the truly differentially genes at 1.3 fold.
Correlation between methylation/gene expression and methylation/copy number
To investigate the effect of DNA methylation on gene expression, we used Spearman's rank to test correlation between the beta-value of a methylation locus and the log2-transformed normalized expression value of a gene within a maximum distance of 2 Kb from that methylation locus. Correlated methylation and expression data were taken from the same samples. In the same way, to see the effect of copy number on gene expression, we also used Spearman's rank to test correlation between the average copy number of a genomic segment and the log2-transformed normalized expression value of a gene overlapping with that genomic segment. Copy number and methylation data were also from the same samples.
Results
Our study was conducted on 24 patients (17 male, 7 female) with CRC with a mean age of 45.5 years (SD 16.8) (Additional File 1 Table S1). There were a total of 27,578 loci covering 14,495 genes (average 1.9 CpG loci per gene) that were studied for methylation status per sample. On average, about 27,511 loci (95% CI 27,488 - 27,534) were detected in each sample at p < 0.05 level. A locus was said to be detected at p < 0.05 level if the mean signal intensity from multiple probes for that CpG locus was significantly higher (at the level of p < 0.05) than the negative control on the same chip. Mean number of loci detected at p < 0.05 in tumor and normal tissue was similar: 27483.88 (SD 114.75) vs. 27507.25 (SD 65.84) (p = 0.41). There was a very strong correlation (r2 = 0.9932) of the total signal intensity (methylated and unmethylated) of the 27,578 loci between the 24 normal tissues and corresponding 24 tumor tissues suggesting uniform amplification and hybridization for all samples. However, when the average β of tumor tissue samples were plotted against that of corresponding normal tissue samples, there were clearly a number of loci that were differentially methylated in CRC tissues compared to normal tissues. The data discussed in the publication have been deposited in NCBI's Gene Expression Omnibus [23] and will be accessible through GEO Series accession number GSE29490 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE29490 webcite).
Sources of variation in the methylation data
Principal component analysis (PCA) suggested a clustering of samples by tissue type (not shown). In the next step, to further investigate the source of variation in the expression, we used multivariate ANOVA. Figure 1A shows the significance of different sources of variation in the entire data in ANOVA model where tissue type (tumor/normal), person-to-person variation (case ID#), sex and location were entered as explanatory variables at a time for the β-value (representing methylation status). The figure shows that "tissue type" and sex were the most significant sources of variation.
Figure 1. A and B Sources of variation in methylation data. Statistical significance of the different sources of variation in the methylation data estimated by 4-way ANOVA models. F-ratio for each factor (source) represents the F-statistics for that factor/F-statistics for error (noise). 1A shows the result in all 27,578 loci and 1B shows similar result only in the autosomal loci (n = 26,486) depicting the effect of sex chromosomal markers.
Differential methylation of colon tissue in male and female subjects
When we compared all 27,578 loci in females with those in males, there were a total of 568 loci differentially methylated at FDR 0.01 level. Interestingly, 551 of them (97.0%) were in the X-chromosome, 2 were in the Y-chromosome and 15 were distributed in the autosomes. This finding is explained by the X-inactivation process, in which one of the two copies of genes on the X chromosome in females is silenced. Considering this fact, we excluded all the sex chromosome markers (n = 1092, of which 1085 in X-chromosome and 7 in Y-chromosome) from subsequent analysis for differential methylation in CRC compared to normal. The significance of different sources of variation in the methylation data in autosomes only is shown in Figure 1B.
Differential methylation in colorectal carcinoma tissue compared to adjacent normal colon tissue
In a total set of 48 samples (tumor and corresponding adjacent normal tissue from 24 patients with CRC), we looked at genome-wide differential methylation in CRC tissue compared to normal tissue. There were a total of 26,486 CpG loci in the autosomes covering 13,890 genes. Here we present the analysis of β-value calculated from background normalized signal intensity. No further normalization of derived β-value was used. In the methylation module of BeadStudio, using the in-built Illumina custom model, we found a total of 875 significantly differentially methylated autosomal loci in CRC tissue compared to normal colonic tissue, of which 275 were hypomethylated (DiffScore = < -30 and delta-β = <-0.2) and 600 were hypermethylated (DiffScore > = 30 and delta β> = 0.2). Univariate and unpaired analysis was used for this.
In the next step, we used multi-way mixed model ANOVA to identify differentially methylated loci in CRC after adjustment for sex, "person to person variation" and location of the tumor (proximal colon or distal colon). To be conservative, we report only differentially methylated loci with absolute delta β of at least 0.2 at FDR 0.01. Following this criteria, we found a total of 852 differentially methylated loci covering 691 genes (see the List_BGN_Auto in the Venn diagram, Figure 2). Figure 2 also shows the lists of differentially methylated loci found using the same criteria (absolute delta β 0.2 at FDR 0.01) for the data with different normalization procedures (as described in statistical method section). There was a good overlap between the normalization procedures, and 627 loci were common to all the analyses. In other words, irrespective of normalization methods (with or without background normalization for signal intensity, with or without quantile normalization of the calculated β-value) these 627 loci were differentially methylated in CRC tissue compared to corresponding normal colonic tissue even after adjustment for sex, person-to-person variation and location of the tumor. Unsupervised clustering based on the common differentially methylated loci divided the samples into two main clusters, and most of the CRC samples were clustered together (figure not shown).
Figure 2. Overlap of results from different normalization methods. Venn diagram showing overlap between the lists of differentially methylated autosomal loci identified by ANOVA models from four different normalization procedures- (a) no normalization of signal intensity, no normalization of β-values- right lower ellipse (n = 724) (b) no normalization of signal intensity to calculate β-values initially, but subsequently quantile normalization of β-values- right upper ellipse (n = 632); (c) background normalization of signal intensity to calculate β-values, but no normalization of β-values- left upper ellipse (n = 852) and (d) background normalization of signal intensity to calculate β-values, and quantile normalization of β-values- left lower ellipse (n = 759).
For further analysis, we focused on these 627 common differentially methylated loci, of which 479 CpG loci were hypermethylated (median distance from TSS 219 bp) and 148 were hypomethylated (median distance from TSS 308 bp) in CRC. Hypomethylated loci were slightly more distal from TSS (Mann-Whitney U test, p < 0.001). On average, 42.8% of the variation in the methylation status of these 627 CpG loci could be explained by tissue (tumor or normal), 26.6% of the variation could be explained by person-to-person variation, 4.52% of the variation was due to sex, 1.42% was due to location and 24.66% of the variation could not be explained by the ANOVA model. Within this list of 627 loci, if we look at the greater magnitude of differential methylation (Delta β = <-0.45 or > = 0.45) or the variation of that loci explained by the tissue type (at least 65%), there were 20 loci covering 17 genes (see Figures 3 and 4). The hypermethylated genes include FLJ25477, ITGA4, DAB2IP, KCNQ5, ZNF625, C1orf165, PRKAR1B, MDFI, C2orf32, RYR2, FLI1, RIC3, TRH, VGCNL1, EYA4 (for q-PCR validation we selected the genes from this list) and the hypomethylated genes include IL21R and PI3. It may be noted that DAB2IP is a known tumor suppressor gene that has been reported to be associated with other cancers [9,24,25]; ITGA4 is also reported to be required for lymphangiogenesis & tumor metastasis [26]. The β-values of some of these loci in CRC and corresponding normal tissue are shown in Figure 5.
Figure 3. Volcano plot of 627 significantly differentially methylated loci. For the 627 DML, the magnitude of differential methylation (Delta β) is shown in the x-axis and the variability of β value explained by the tissue type (CRC or normal) is shown on the y-axis. Color coding was done by the distance of the locus from the transcription start site (TSS), where blue indicates close & red indicates away from the TSS.
Figure 4. Heatmap of 20 highly differentially methylated loci. Unsupervised hierarchical clustering of 20 DML (rows) in 48 samples (columns) that are shown in Figure 3. These 20 DML represent the most highly differentially methylated loci (Δ β ≤ -0.45 or ≥ 0.45 with FDR 0.01) and/or whose variation could be explained by tissue type (at least 65%) in our ANOVA models. Location of the tumor (proximal or distal colon) and the differentiation of the tumor (moderately differentiated or poorly differentiated) are shown above the heatmap.
Figure 5. Dotplot showing differential methylation. The β-values for four genes (DAB2I, ITGA4, PRKAR1B and IL21R) in both normal (blue) and CRC (red) tissue samples. Connecting lines indicate paired samples (indicating same patient). DAB2I, ITGA4 and PRKAR1B show hypermethylation in the CRC samples when compared to the normal tissue and IL21R shows hypomethylation in the CRC samples when compared to the normal tissue.
In fact, there was also a very good overlap between the univariate analysis of BeadStudio Methylation module and the mixed model multi way ANOVA analysis. A total of 832 loci were common between the lists generated by univariate Illumina custom model (n = 875) and the list generated by mixed model multi way ANOVA analysis (n = 852) as mentioned above.
If we looked at the multiple loci near a single gene, usually all the loci showed a similar trend. For example, there were six loci for the gene ESR1, and all were hypermethylated in CRC. We also took the average β values of multiple markers from the same gene and looked for differentially methylated genes in CRC (not shown here), and the result was almost similar to what we see with probe level analysis.
GO Enrichment Analyses of the lists of differentially methylated genes in colorectal carcinoma tissue compared to adjacent normal colon tissue
We examined the list of 627 loci covering 513 genes (479 loci representing 374 genes were hypermethylated and 148 loci representing 139 genes were hypomethylated) to see if any particular group of genes were found to be differentially expressed in the ANOVA models more frequently than by chance. The Gene Ontology (GO) database (http://www.geneontology.org webcite) categorizes genes on the basis of (a) "molecular function", (b) "biological process" and (c) "cellular component". For example, the number of hypermethylated loci (n = 479) in the list represent only 1.8% of the total autosomal loci (n = 26486). In other words, if 479 loci were picked randomly, then we would not expect more than 1.8% of the loci from any particular category to be present in that list. GO-enrichment analysis tests if a group of genes is overrepresented (i.e., enriched) in a list than would occur by chance. The higher the Enrichment Score (ES), the more significant the enrichment is. This is used for ranking the groups. For the list of hypermethylated loci, if we use "molecular function" for categorization, then we see that the top-ranking groups of genes that were enriched include "transmembrane receptor activity", "receptor activity", "transcription factor activity" "G-protein coupled receptor activity", "ionotropic glutamate receptor activity", "glutamate receptor activity", "extracellular-glutamate-gated ion channel activity" and "transmembrane receptor protein phosphatase activity". In other words, most of these groups or subgroups are under the broader categories of "receptor activity" or "transcription factor activity" (Figure 6). Similarly, if we use "biological process" for categorization, then the genes under the broader category "cellular developmental process" and its sub-class "cell differentiation" were highly enriched in the list of hypermethylated loci.
Figure 6. GO-Enrichment Analysis of the list of hypermethylated loci. Enrichment score (ES) of subgroups under "Transmembrane receptor activity" which was the most enriched (ES = 30) among the "molecular function" category.
Differential methylation profile at "gene set" level in colorectal carcinoma
After looking into the differential methylation at the individual gene level, we also looked for differential methylation of different "gene sets" (different groups of genes) in colorectal carcinoma by using Gene Set Enrichment Analysis (GSEA) as well as GO-ANOVA. Gene sets were defined using publicly available data from the GO website (http://www.geneontology.org webcite). GSEA revealed that a total of 512 "gene sets" were differentially methylated as the normalized enrichment score (NES) was = <-1.5 (n = 270, hypomethylated) or NES > = 1.5 (n = 242, hypermethylated). Using the permutation p-value for the Enrichment Score (ES) as a cut-off, a total of 220 "gene sets" were differentially methylated at p = <0.01. In GO-ANOVA analysis a total of 2851 "gene sets" crossed the threshold of FDR 0.01, and a total of 932 gene sets showed an average delta-β of = <-0.1 (n = 384, hypomethylated) or > = 0.1 (n = 548, hypermethylated). Additional File 1 Table S2 shows the results from GSEA as well as GO-ANOVA for the gene sets that had p = <0.01 for ES, NES either = <-1.5 or > = 1.5 in GSEA and also showed significant GO-ANOVA p-value at FDR 0.01 and an average delta β = <-0.1 or > = 0.1. The gene sets are arranged by NES in descending order in Additional File 1 Table S2. Obviously sorting by GO-ANOVA p-value would change the ranking. The major groups or "gene sets" that are hypermethylated in CRC are "inhibition of adenylate cyclase activity by G-protein signaling" (Figure 7), "Rac guanyl-nucleotide exchange factor activity", "regulation of retinoic acid receptor signaling pathway" and "estrogen receptor activity".
Figure 7. GO-ANOVA for a "gene set". GO-ANOVA result for differential methylation of the gene set "inhibition of adenylate cyclase activity by G-protein signaling". The markers in this "gene set" are shown in the x-axis and their corresponding β-value is presented on y-axis. Error bars represent SE.
Comparisons to colorectal carcinoma signatures from prior studies
We compared our conservative list of 513 differentially methylated genes to lists obtained by a number of previous studies. In a 2010 review paper, Kim et al. [27] compiled a comprehensive list of differentially methylated genes in CRC tissue and other clinical samples from patients with CRC. These were mainly candidate gene approach based studies. There were a total of 59 unique genes reported to have differential methylation in CRC tissue. Out of those 59 genes, 40 of them were also studied in our present study. In fact there were 245 loci on the chip we used that covered these 40 genes. It may be noted that 17 of those 40 genes (42.5%) were also found in our conservative list of 513 differentially methylated genes. In addition to comparing the gene lists, we also looked at the differential methylation pattern of these 245 loci covering those 40 genes in our setting. A volcano plot (Additional File 3 Figure S2A) clearly shows that most of these loci were differentially methylated, but the number actually depends on the strictness of the criteria used to define differential methylation. In fact 32 of them (80%) were significant at FDR 0.05 level without considering the delta β. Recently Ang et al. published a study on CRC using a genome wide approach using Illumina's GoldenGate methylation panel of 1505 CpG loci [9]. They reported a total of 202 differentially methylated loci in CRC covering 132 genes [9]. Of those 132 genes, 37 were common to our list. But again, if we look at the methylation data, then we see that in our assay there were a total of 376 loci covering those genes and 263 of those loci (70%) were also differentially methylated at FDR 0.05 in our data set (Additional File 3 Figure S2B). Therefore, it is important to take the selection criteria of a list into account while comparing the gene list. Hence, our genome-wide methylation assay not only clearly supports most of the previous findings from the literature, but in addition to that we found a large number of novel differentially methylated loci in CRC tissue compared to surrounding healthy colon tissue. The complete list of differentially methylated loci is presented as additional material (Additional File 1 Table S3) that shows the loci that have been previously reported as well as the novel loci found in present study. Kim et al. [27] also compiled a list of genes that were reported in the literature to be differentially methylated in other clinical samples (serum/plasma or stool) from CRC patients. The authors reported a total of 19 unique genes (there were 129 loci in the chip we used that covered 18 of these 19 genes) and interestingly 9 (50%) of those genes were also found in our conservative list of 513 differentially methylated genes in CRC. In fact, 63 of the loci covering 14 of those 18 genes (77.8%) were significantly differentially methylated at FDR 0.05 level.
Additional File 3. Figure S2: Volcano plot showing methylation status of previously reported genes in our samples. The Delta β is shown on x-axis and ANOVA p-value on the y-axis. A: represents the 245 loci covering the previously reported genes mainly from candidate gene approach-based studies; B: represents the 376 loci covering the 132 genes reported from a single study based on genome-wide approach (although testing only 1505 CpG sites). The side bar shows the color scale depending on Delta β where blue indicates hypomethylation and red indicates hypermethylation in CRC.
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In addition to looking only at CRC methylation signatures in the literature, we also compared our list to that of hypo- and hyper-methylated genes in cancer as a whole. In a 2009 review, Pogribny et al. compiled a list of 38 unique genes that are reported to be hypomethylated in different human cancers [28]. Only four of those (ESR1, HSPE2, TCL1 and TNFRSF8) were common in our list, and we found that all four of them were found to be hypermethylated to some degree in CRC in our study. In contrast these four genes were reported to be hypomethylated in different cancers - endometrial carcinoma, prostate cancer, T-cell Lymphocytic leukemia and in Hodgkin lymphoma respectively.
In a2007 review, Esteller et al. [29] compiled a list of 47 genes that were reported to be hypermethylated in different human cancers. Eight of those (CDH13, CDKN2A, ESR1, TMEFF2, GATA4, SFRP1, TP73 and SOCS3) were common in our list and all were hypermethylated in CRC in our study.
DNA repair genes are known to be important for the pathogenesis of carcinoma in general. In a review, Ronen and Glickman compiled a list of 261 DNA repair genes [30]. It was interesting to note that none of these 261 genes were common to our list of 513 genes with differential methylation in CRC.
Uses of methylation signatures
We attempted to identify some models for differentiating CRC samples from normal samples based on methylation status. The models were identified using a 2-level nested cross validation method. Though an independent sample set was unavailable for this study, this method provided a means to estimate the accuracy of the models that may be expected in an independent set of samples. However we agree that statistical model can not replace the need of validation in an independent set of real samples.
The overall idea was to set aside a random set of samples, and then use the rest of the samples from the present study to identify an optimal combination of loci that would classify the samples as CRC or normal, and then to test the expected accuracy and different test characteristics [sensitivity, specificity, positive predictive value and negative predictive value] of the model in a different set of samples that was held out initially. Table 1 shows the summary of different models. For example, the model with 4 loci - Illumina ID# cg02656594, cg13577076, cg20415809 and cg27650175 is expected to correctly predict the diagnosis (normal or CRC) in 94% of the samples. These loci are located close to the transcription start site of the genes IL21R, PRKAR1B, ITGA4 and DAB2IP respectively. It may be noted that the other models involving more variables also give reasonable sensitivity (around 89%), specificity (around 95%), positive predictive value (around 95%) and negative predictive value (around 89%). Therefore, these markers may be considered for clinical application.
Table 1. Results from two-level nested cross-validation
We also attempted to identify methylation signatures that could differentiate histopathological findings in CRC. Although the present study was not designed to address this issue, we analyzed the 24 CRC samples using different phenotypes, including tumor stage, tumor grade, differentiation of the tumor, tumor infiltration by lymphocytes, extracellular mucin and signet ring cell. Irrespective of the histopathological diagnosis (adenocarcinoma or mucinous adenocarcinoma) and age at diagnosis (= <45 yrs vs. >45 yrs), a total of 14 loci were significantly differentially methylated in CRC of the proximal colon than those of the distal colon. Unsupervised hierarchical clustering of those loci in the CRC samples is shown in the Additional File 4 Figure S3. GO-Enrichment analysis of these 14 genes showed significant enrichment of genes related to "gut morphogenesis".
Additional File 4. Figure S3: Heatmap of 14 loci that are differentially methylated in proximal CRC compared to distal CRC. Unsupervised hierarchical clustering of the 14 loci (rows) in 24 CRC samples (columns). Thirteen of these loci were hypermethylated in proximal CRC compared to distal CRC. The two major clusters generated (top dendogram) in this analysis separated most of the proximal CRC tissues from the distal CRC tissues. Age at diagnosis (>45 yrs or = <45 yrs and the differentiation of the tumor (moderately differentiated or poorly differentiated) are shown above the heatmap.
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Results from q-PCR validation
We selected the top-ranking hypermethylated 12 genes (shown in Figure 3) for validation of the Infinium methylation platform data using a methyl profiler assay. Assay development was not feasible for PRKAR1B for technical reasons and so that gene was replaced by the next highest-ranking gene, TRH. For validation we used paired DNA samples from 10 randomly selected patients from the same sample set. To compare the data from q-PCR to the β-value in the microarray, we added the proportion of intermediate methylation and hypermethylation in the q-PCR data to obtain the proportion of methylated DNA. Figure 8 and Additional File 1 Table S4 summarizes the differential methylation of these genes in CRC tissue compared to corresponding adjacent normal colonic mucosa. It may be noted that 10 out of these 12 genes were also found to be significantly hypermethylated in the q-PCR experiment. Among these 10 validated genes, 7 are novel (TRH, C2orf32, FLJ25477, KCNQ5, C1orf165, MDFI and RIC3) and the remaining three (ITGA4, DAB2IP and FLI1) were previously reported by others [9,27]. The correlation coefficients ("r") also suggest reasonably good correlation between the q-PCR data and the microarray data (Additional File 1 Table S4).
Figure 8. Validation of methylation by q-PCR. Differential methylation of selected genes in CRC tissue (_CRC) compared to adjacent normal colon mucosa (_N), as determined by Methyl Profiler q-PCR assay.
Does the differential methylation status correlate with chromosomal abnormalities and differential gene expression in CRC?
To address this issue, we also did a high density oligonucleotide SNP array (610 Quad) to detect cytogenomic abnormalities and a genome-wide gene expression assay (HT12 v4), for the same 24 patients. We detected a total of 1196 genomic segmentation regions (harboring 970 genes), for which the copy number significantly (p < 0.05, chi-square test) differs between CRC and normal mucosa. We also detected a total of 1399 down-regulated and 1209 up-regulated genes (at least 1.3 fold at FDR 0.01) in CRC compared to normal mucosa. Only 28 hypermethylated genes were down-regulated, and 48 down-regulated genes were among the genes located in genomic regions that show significant copy number change in CRC compared to normal colonic mucosa (Figure 9A). Similarly, only 6 of the hypomethylated genes were up-regulated in CRC, and 60 up-regulated genes were among the genes located in genomic regions that show significant copy number change in CRC compared to normal colonic mucosa (Figure 9B). In other words, there are relatively few genes for which either differential methylation or copy number change alone can account for the observed changes in gene expression. This clearly depicts the complexity of genomic and epi-genomic interplay in carcinogenesis. Figure 9C shows the heatmap for gene expression of those 25 genes which are down-regulated by hypermethylation irrespective of CN status.
Figure 9. A Overlap of hypermethylation, down-regulation and differential copy number state. B: Overlap of hypomethylation, up-regulation and differential copy number state. C: Heatmap of differentially expressed genes: Hierarchical clustering of the gene expression data from the 25 hypermethylated genes (rows) shown in Figure 9A across the samples (columns). These genes do not overlap with regions showing significantly different copy number state in CRC tissue. The clusters generated (top dendogram) in this analysis separated most of the CRC tissues from the normal colonic mucosa tissues.
In general, statistically significant cis-correlation (with in 2 kb region, with rank correlation p = <0.05) between methylation and gene expression was observed at 704 loci. However, only a few of these genes were differentially methylated or expressed in CRC compared to normal tissue (Figure 10A as example), while for many of the genes the methylation status correlated with gene expression at sample level without being differentially methylated or expressed in CRC tissues (Figure 10B). In the same line, we found that in a total of 3850 genomic segmentation regions, the gene expression was significantly correlated (rank correlation p = <0.05) to copy number status of the region harboring the gene. But only a few of these genes were differentially expressed or showed differential CN change in CRC compared to normal tissue (Figure 10C as example), while for many of the genes the expression level correlated with genomic CN status at sample level without being differentially expressed or having differential CN status in CRC tissues (Figure 10D as example).
Figure 10. Correlation of gene expression with methylation and copy number data. A and B: Examples of gene expression (y-axis) correlating to methylation (x-axis) with differential expression & methylation in CRC compared to normal (A) and without differential expression or methylation (B). C and D: examples of gene expression (y-axis) correlating to copy number (x-axis) with differential expression & CN change in CRC compared to normal (C) and without differential expression or CN change (D).
Discussion
There are only a few studies addressing the genome-wide methylation in colorectal carcinoma [9,31-33]. Very recently Kim et al [32] (Feb 2011) and Oster et al [33] (Mar 2011) used the same commercially available Infinium methylation 27 arrays in CRC and identified differentially methylated sites in Korean and European population respectively. Oster's study used carcinoma and normal tissue from different individuals for methylation analysis, whereas ours used paired tumor and adjacent normal tissue from the same patient. This allowed us to eliminate inter-individual variation in our methylation analysis, which may be one reason why our study detected a larger number of differentially methylated genes in carcinoma. Kim's study compared the methylation in paired samples like ours, but they looked at gene expression in a different set of individuals. While they looked for effect of methylation on gene expression, they could not find statistically significant difference in the mRNA expression level between promoter hypermethylation group and hypomethylation group, whereas we were able to calculate correlation coefficients using paired data in every gene and found several significant correlations. The only other genome-wide methylation study in CRC addressing promoter CpG loci using commercially available array, used much lower density array (only 1505 CpG loci) [9].
Our study in south-east Asian population suggests that, in comparison to the normal colonic mucosa, the corresponding CRC tissue shows a large number of differentially methylated loci within the CpG islands close to the transcript start site of genes, indicating the role of DNA methylation in the pathogenesis of colorectal carcinoma. The results from our study not only confirm the findings from many previous candidate gene approach based studies, but we also report a large number of novel loci that show differential methylation in CRC.
We noticed the influence of sex on genome-wide methylation that is explained by the X-inactivation process, in which one of the two copies of genes on the X-chromosome in females is silenced. A similar finding was also recently reported by Liu et al. [34].
Laird et al. [35] has recently focused on the different statistical issues for methylation data. We applied different normalization methods and found considerable overlap between the results. Use of stringent criteria for selecting differentially methylated loci and the considerable overlap between the results from different analyses, the 2-level cross validation and finally the q-PCR validation in subset suggest that we detected the truly differentially methylated loci in CRC.
Recently Irizarry et al. used a Comprehensive High-throughput Array for Relative Methylation (CHARM) assay to show that most methylation alterations in CRC occur up to 2000 bp away from the CpG islands themselves [31]. Because of the design of the chip used in the present study, we did not have the opportunity to look at the differential methylation at loci > 1500 bp away from the TSS. However, similar to results from Irizarry et al. [31], we also found that the hypomethylated loci were slightly more upstream than the hypermethylated loci.
The cross-validation results are very encouraging as a potential biomarker, but we have cross-validated only in colon tissues and not in circulating plasma DNA. In the future we would like to test the markers in an independent sample set of circulating plasma or serum DNA in CRC patients and healthy individuals. Recently He et al. [36] selected three methylation markers from the published literature and tested the practical use of those markers in peripheral blood sample from CRC patients. They found a sensitivity of 81% and a specificity of 90%. We had the advantage of profiling a very large number of CpG loci in paired CRC and normal colonic mucosa tissue, and our 2-level cross validation suggested that the four markers could be used as biomarkers with slightly better test characteristics.
Tanaka et al. [37] have recently applied an analytical strategy known as structural equation modeling to understanding methylation in CRC. Using a large database of over 800 samples, the authors were able to construct causality pathways of KRAS and BRAF mutations, as well as various phenotypes, on methylation of specific genes. This strategy was not feasible for our current study because of our smaller sample size and because we had not obtained information on KRAS and BRAF mutations. Nonetheless, it will be valuable for our planned future study with an expanded cohort.
Illumina's methylation assay has been compared to other platforms by others and has shown dependable results with the correlation ranging from 0.8 to 0.9 [32,33,38]. We also have validated the methylation data form Infinium methylation for 12 of the highly differentially methylated genes in our study and also found similar high correlations with Methyl Profiler assay (see Additional File 1 Table-S4 and Additional File 5 Figure S4). In another study, reproducibility tests of Infinium methylation platform was reported to have correlation greater than 0.98 between technical replicates [39]. We are aware of the fact that Illumina's Methylation27 assay detects the methylation status of on average ~2 CpG sites per gene for most genes. However, for the genes for which there were multiple CpG loci on the array (e.g. ESR1 or DAB2IP), we found all of the loci to be differentially methylated in the same direction. We also validated Illumina's platform in the top-ranking genes by methyl profiler PCR array which is (a) not dependent on bisulfite conversion and also (b) provides an overall methylation status of the target region as opposed to single loci. This paper was focused mainly to look at DML in CRC. However, we have also explored the link between chromosomal abnormalities (copy number), methylation and gene expression. Regulation of gene expression is complex and is not dependant only on methylation status or copy number status. Using integration of molecular cytogenetics, genome-wide copy number and expression microarray profiling, Camps et al have demonstrated the effect of copy number on gene expression in CRC [40]. To our knowledge, our study is the first one to comprehensively look at the genome-wide methylation, copy number and gene expression - all three together in primary CRC tissue. In our study, expression of a small proportion of genes was found to be correlated to methylation and another small proportion was correlated to copy number changes seen in CRC. Although methylation status of many loci could not explain the functional relevance to gene expression, these promoters methylation may be used clinically as biomarkers.
Additional File 5. Figure S4. Comparison between q-PCR and microarray methylation data. Graphs are shown for the 12 genes validated by q-PCR. The y-axis plots the β value from microarray data. The x-axis plots the proportion of intermediately methylated and hypermethylated DNA.
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Conclusions
Our genome-wide methylation study in CRC clearly supports most of the previous findings from the literature, and in addition to that we found a large number of novel DML in CRC tissue, some of which may be used for clinical application. Further study is warranted to confirm these findings.
Authors' contributions
MGK conceived and designed the study, performed data analysis and wrote the manuscript, FJ designed and carried out the genome-wide methylation assay and drafted the manuscript, MR collected the tissue samples and did the histopathology, SR processed the tissue samples and carried out the gene expression and validation assay, RPB processed the tissue samples, and helped in methylation microarray and high density SNP array; RR and CD helped in manuscript, MRZ helped in sample collection and transportation of the samples to USA, MK organized & supervised the tissue collection and was responsible for histopathology, HA helped in manuscript, supported and coordinated the study. All authors read and approved the final manuscript.
Acknowledgements
This work was supported by the National Institutes of Health grants U01 CA122171, P30 CA 014599, P42ES010349, R01CA102484, and R01CA107431.
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Pre-publication history
The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1755-8794/4/50/prepub
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Recycling as Habitual Behavior: The Impact of Habit on Household Waste Recycling Behavior in Thailand
Achapan Ittiravivongs
Abstract
This research aims to permit a better understanding of factors influencing recycling behavior of Thai households in a habitual perspective. The study applied theory of interpersonal as critical framework and investigated the role of habit on recycling involvement of 381 samples in Bangkok. The outcomes indicated that recycling behavior was significantly predicted by recycling intention, habit, recycling ability, facility condition, and adequacy of recycling information, in order of strength. A trade-off relationship between recycling habit and intention was also found. With higher degree of habit, recycling behavior is subjected to be less depended on intention. In addition, relations of behavior-intention and behavior-facility condition were found significantly different across habit levels. Recycling behavior is likely to be less related to recycling intention and facility condition for strong habit group. The results suggested that recycling habit is an important issue needed to be considered as a notable factor influencing household recycling behavior.
Full Text: PDF DOI: 10.5539/ass.v8n6p74
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Asian Social Science ISSN 1911-2017 (Print) ISSN 1911-2025 (Online)
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Hadeland, Land og ValdresEdit This Page
From FamilySearch Wiki
Probate records for Hadeland, Land og Valdres, 1659-1857 judicial districts in Oppland county. They include these places:
• Lunner
• Jevnaker
• Gran
• Søndre Land
• Nordre Land
• Etnedal
• Sør-Aurdal
• Nord-Aurdal
• Vestre Slidre
• Østre Slidre
• Vang
For later probate records see the combined Hadeland og Land judicial districts and Valdres judicial district.
The probate records usually contain the date of probate, name of the deceased, names of heirs, guardians and trustees, inventory lists, land and holdings, and other miscellaneous information.
Return to Oppland County
Return to Norway
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• This page was last modified on 7 January 2011, at 07:35.
• This page has been accessed 426 times.
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Sonoma County, CaliforniaEdit This Page
From FamilySearch Wiki
Revision as of 17:06, 10 March 2009 by Grobie (Talk | contribs)
United States > California > Sonoma County
Contents
County Courthouse
History
Parent County
1850--Sonoma County was created 18 February 1850 as an original county. County seat: Santa Rosa [1]
Boundary Changes
Record Loss
1906 -- Courthouse burned and many records were damaged.
For further information on researching in burned counties, see the following:
Places/Localities
Populated Places
Neighboring Counties
Resources
Cemeteries
Church
LDS Ward and Branch Records
• Santa Rosa
Court
Land
Local Histories
Maps
Military
Newspapers
Probate
Taxation
Vital Records
Societies and Libraries
Sonoma County History & Genealogy Library
Web Sites
• USGenWeb project. May have maps, name indexes, history or other information for this county. Select the state, then the county.
• Family History Library Catalog
References
1. The Handybook for Genealogists: United States of America,10th ed. (Draper, UT:Everton Publishers, 2002).
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About this Journal Submit a Manuscript Table of Contents
Evidence-Based Complementary and Alternative Medicine
Volume 2012 (2012), Article ID 673132, 7 pages
doi:10.1155/2012/673132
Research Article
Effects of Berberine and Hwangryunhaedok-Tang on Oral Bioavailability and Pharmacokinetics of Ciprofloxacin in Rats
1KM-Based Herbal Drug Research Group, Korea Institute of Oriental Medicine, Daejeon 305-811, Republic of Korea
2Laboratory of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Chungnam National University, Republic of Korea
Received 31 July 2012; Revised 25 September 2012; Accepted 26 September 2012
Academic Editor: Bhushan Patwardhan
Copyright © 2012 Youn-Hwan Hwang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Linked References
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22. P. Joyanes, A. Pascual, M. J. Giménez, I. García, L. Aguilar, and E. Perea, “Differences between two new quinolones (gemifloxacin and trovafloxacin) and ciprofloxacin in their concentration-dependent killing of Streptococcus pneumoniae,” Chemotherapy, vol. 47, no. 6, pp. 409–414, 2001. View at Publisher · View at Google Scholar · View at Scopus
23. C. E. Thorburn and D. I. Edwards, “The effect of pharmacokinetics on the bactericidal activity of ciprofloxacin and sparfloxacin against Streptococcus pneumoniae and the emergence of resistance,” Journal of Antimicrobial Chemotherapy, vol. 48, no. 1, pp. 15–22, 2001. View at Scopus
24. E. Szałek, A. Kamińska, J. Gozdzik-Spychalska, E. Grześkowiak, and H. Batura-Gabryel, “The PK/PD index (CMAX/MIC) for ciprofloxacin in patients with cystic fibrosis,” Acta Poloniae Pharmaceutica, vol. 68, no. 5, pp. 777–783, 2011. View at Scopus
25. M. E. Cavet, M. West, and N. L. Simmons, “Fluoroquinolone (ciprofloxacin) secretion by human intestinal epithelial (Caco-2) cells,” British Journal of Pharmacology, vol. 121, no. 8, pp. 1567–1578, 1997. View at Publisher · View at Google Scholar · View at Scopus
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About this Journal Submit a Manuscript Table of Contents
International Journal of Pediatrics
Volume 2012 (2012), Article ID 478610, 6 pages
doi:10.1155/2012/478610
Research Article
Adolescent Metabolic Syndrome Risk Is Increased with Higher Infancy Weight Gain and Decreased with Longer Breast Feeding
1Division of Child Development and Community Health, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0927, USA
2Institute of Nutrition and Food Technology (INTA), University of Chile, El Líbano 5524, 138-11 Santiago, Chile
3Center for Human Growth and Development and Department of Pediatrics and Communicable Diseases, University of Michigan, 300 North Ingalls, 10th Floor, Ann Arbor, MI 48109-5406, USA
Received 15 December 2011; Revised 1 March 2012; Accepted 18 May 2012
Academic Editor: Ricardo D. Uauy
Copyright © 2012 Kim Khuc et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background. Prevalence of the metabolic syndrome is increasing in pediatric age groups worldwide. Meeting the criteria for the metabolic syndrome puts children at risk for later cardiovascular and metabolic disease. Methods. Using linear regression, we examined the association between infant weight gain from birth to 3 months and risk for the metabolic syndrome among 16- to 17-year-old Chilean adolescents (), accounting for the extent of breastfeeding in infancy and known covariates including gender, birth weight, and socioeconomic status. Results. Participants were approximately half male (51%), born at 40 weeks of gestation weighing 3.5 kg, and 48% were exclusively breastfed for days. Factors independently associated with increased risk of metabolic syndrome in adolescence were faster weight gain in the first 3 months of life (, ) and male gender (, ). Breastfeeding as the sole source of milk for days was associated with significantly decreased risk of metabolic syndrome (). Conclusion. This study adds to current knowledge about early infant growth and breastfeeding and their long-term health effects.
1. Introduction
As the prevalence of obesity in children and adolescents has escalated worldwide, signs of the metabolic syndrome (MetS) are increasingly observed in the pediatric age range [1]. MetS refers to a cluster of abnormal physical examination and laboratory findings, including high waist circumference, serum triglyceride, serum glucose, and blood pressure, and low-serum HDL-cholesterol. These findings synergistically relate to risk for developing type 2 diabetes and cardiovascular diseases (CVDs), including coronary artery disease and stroke [24]. In a study of a representative sample of US 12- to 19-year-old, 8.6% met criteria for MetS; Hispanic youth had a higher prevalence (11.2%) than white (8.9%) or black adolescents (4.0%) [2]. Children who meet the criteria are at increased risk for CVD in adulthood [5]. Focusing on the MetS during the pediatric period is expected to lead to early-prevention strategies for diabetes and CVD [5].
A large body of evidence suggests that metabolic programming can occur early in life [68]. Early-life risk factors include low birth weight and rapid postnatal weight gain. Breastfeeding including duration and dose appears to offer protection for obesity, type 2 diabetes, the MetS, and CVD [912]. In fact, the time immediately before and after birth may be a sensitive period related to metabolic and cardiovascular risk [13]. Rapid post-natal weight gain is associated with increased risk for obesity, type 2 diabetes and hypertension in young adulthood [1318]. Infant weight gain, especially in the first 3 months, may be more important than birth weight as a predictor of later health outcomes [13]. Adolescent MetS has previously been found to be associated with infancy growth in the setting of a developed country [19].
Much of the work on fetal origins of disease has been done in developed countries beginning with Barker’s work in England [7]. Research in low- to middle-income countries is needed to further delineate the roles of biology and environment related to early-life risk for cardiovascular disease and related conditions. Our cohort of low- to middle-income Chilean adolescents, studied since infancy, provides a special opportunity to address these research questions, especially because the participants were born during a period of rapid nutritional and economic transition in Chile. This transition was characterized by economic progress that led to increased consumption of calories, fat, animal protein, and processed foods, and increased mortality from noncommunicable chronic diseases [20]. The aims of this study were to examine the association between infant weight gain from birth to 3 months and risk for the MetS in mid-adolescence, accounting for the extent of exclusive breastfeeding in infancy and covariates known to be associated with infant growth and the MetS, gender, birth weight, and socioeconomic status (SES).
2. Methods
2.1. Cohort
This is an observational cohort study involving adolescents who were enrolled as infants in a randomized controlled trial of iron supplementation to prevent iron deficiency anemia. Infants were enrolled from 1991–1996 in Santiago, Chile; 1657 infants completed the preventive trial at 1 year. The inclusion criteria for the preventive trial were infant birth weight of 3 kg or more, with no birth complications, major congenital abnormalities, or prior iron therapy. Due to a highly successful national breastfeeding campaign, all but 8 infants in the cohort were initially breastfed. Infants were randomly assigned to low or high iron supplementation, or usual nutrition (no added iron). A more detailed description of randomization techniques, sampling, and entrance and exclusion criteria is published elsewhere [21]. The participants have been involved in follow-up studies at 5, 10, and 16 years. At 16 years, the participants from the longitudinal cohort were invited to enroll in a study of adolescent obesity and cardiovascular risk. We report on the first 384 studied from a randomly selected sample of the original cohort evaluated at 16 years between May 2009 and January 2011. Complete data from the infancy and adolescent waves were available for 357 of the 384 adolescents. Infancy variables (birth weight, weight at 3 months, and gestational age) did not differ between the 357 studied and the original 1657 infant participants. Our analytic sample was more likely to receive bottle supplementation before 90 days, compared to the larger cohort (52% versus 45%, ). The longitudinal study has been approved by the institutional review boards of the University of Michigan, Ann Arbor USA; Institute of Nutrition and Food Technology, University of Chile, Santiago, Chile for each wave of study; by the University of California, San Diego, for the 16-year study of obesity and cardiovascular risk.
2.2. Infancy Data
We included the following infancy measures: gender, weight measured at birth and at 3 months, and date of the first supplemental bottle. Maternal education was used as a proxy for SES. Mother’s prepregnancy BMI was calculated from measured height and self-report of prepregnancy weight. Data on pre-pregnancy weight was not collected for the infancy study. During the 10-year wave of data collection, mothers reported their pre-pregnancy weight; it was highly correlated with their actual weight 10 years later.
2.3. Adolescent Data
Adolescents were assessed between 16 and 17 years during the fourth wave of the longitudinal research study (infancy, 5 years, 10 years, and 16-17 years). Height (cm), weight (kg), waist and hip circumference (cm), and blood pressure (mm Hg) were measured by a physician-investigator at the nutrition research center. Standardized procedures [22] were used to measure weight to the closest 0.1 kg, using a SECA scale, and height to the closest 0.1 cm, using a Holtain stadiometer. Measurements were taken twice, with a third measurement if the difference between the first two exceeded 0.3 kg for weight and 0.5 cm for height. Fasting serum triglyceride, cholesterol, and glucose levels were performed. Serum glucose concentration (mg/dL), triglycerides (mg/dL), and cholesterol (mg/dL) levels were determined using an enzymatic-colorimetric test (QCA S.A., Amposta, Spain). Using a standardized questionnaire, parents reported family history of type 2 diabetes, hypertension, dyslipidemia, and heart attack before the age of 60, in first-degree relatives.
2.4. Statistical Analysis
Infant weight gain in the first 3 months was calculated as weight gain velocity over the first 3 months (91.3 days): (weight [kg] at 3-month birth weight [kg])/(age at 3-month measurement * 91.3 days). Extent of breastfeeding was assessed as a dichotomous variable representing breastfeeding without bottle supplementation for less than 90 days, compared to 90 days or more. Data was not available on introduction of complementary foods. Maternal education was assessed as a continuous measure (median for sample = 10 years). We constructed a metabolic syndrome risk z-score according to the work of Brage and colleagues [23]. The following variables were converted to z-scores: the reciprocal of the HDL value, the mean of the systolic and diastolic blood pressure measurements, waist circumference, fasting serum triglyceride, and glucose. We obtained a continuous, normally distributed metabolic risk z-score by averaging these 5 values.
For descriptive statistics, continuous variables were expressed as median and interquartile ranges and categorical variables as frequencies. We evaluated cardio/metabolic risk factors and overall prevalence of the MetS according to International Diabetes Federation definition [24]: waist circumference ≥94 cm for boys and ≥80 cm for girls, plus any two of the following four factors: triglycerides ≥1.7 mmol/L, HDL-cholesterol <40 mg/dL in males and <50 mg/dL in females, systolic BP ≥130 or diastolic BP ≥85 mm Hg, fasting plasma glucose ≥100 mg/dL. BMI percentile was described according to CDC standards.
We used SPSS for Windows version 18.0 (Chicago, IL, USA), a P value of <0.05 denoted statistical significance. Multiple linear regression models were used to determine the relationship between change in weight (kg) in the first 3 months and metabolic syndrome risk z-score, adjusting for extent of breastfeeding and the following covariates: birth weight, gender, SES, age, mother’s age at birth of infant, mother’s pre-pregnancy BMI, and family history of type 2 diabetes, dyslipidemia, and heart attack. We tested the full model and then, using backward elimination, removed each variable that was not significantly related to the outcome in the model based on a significant P value of <0.05. As the sample came from an iron-deficient anemia prevention trial, we tested whether iron-deficient anemia during the first year of life or iron supplementation were significant covariates in our models. Neither variable showed significant relationship in the models and were thus removed from the final models.
3. Results
Participants were assessed at a mean age of 16.6 years. Males and females represented about 51% and 49% of the sample, respectively. Participants had been born at 40 weeks of gestation weighing 3.5 kg, on average, and 48% were exclusively breastfed for ≥90 days. The median BMI percentile was 68.7 with 15.2% in the obese range and 10.4% met criteria for MetS. Table 1 describes infant and family background characteristics by gender of the 357 participants in infancy and adolescence. Cardio/metabolic risk factors are also described. Males had higher birth length, weight at three months, and higher weight gain between birth and 3 months compared to females. Males also had significantly lower HDL cholesterol and higher blood pressure values (systolic and diastolic), glucose and MetS risk z-scores than females. There were no significant differences between males and females in gestational age, birth weight, maternal education, exclusive breastfeeding for ≥90 days, and prevalence of the MetS.
Table 1: Background characteristics of study participants by gender.
The multiple variable linear regression models (full and final) are shown in Table 2. The final model revealed that weight gain over the first three months was associated with an increased MetS risk score at 16-17 years, taking into account extent of breastfeeding and gender (, 95% , 0.27, ). Introduction of the first bottle at 90 days or after was related to a lower MetS risk score in adolescence (, 95% , −0.04, ), taking into account other covariates. Additionally, being male was associated with an increased MetS risk score in the model (, 95% , 0.37, ). The final model explained 9% of the variance in MetS risk.
Table 2: Linear regression models to determine adjusted associations with adolescent MetS risk ().
4. Discussion
We examined weight gain in the first 3 months of life and timing of bottle supplementation related to MetS risk at 16 years. In both sexes, adolescents who had more rapid weight gain during the first 3 months of infancy had higher adolescent MetS risk scores compared to those who gained less weight in early infancy. The association of weight gain with MetS risk is consistent with findings from a study addressing the same question, in a Scandinavian country [19]. Infancy weight gain has previously been associated with later obesity in childhood and adulthood [15, 18, 25, 26]. In addition, especially for low-birth-weight infants, more rapid early weight gain, sometimes called catch-up growth, has been related to higher risk of developing type 2 diabetes and/or cardiovascular disease [10]. Contrary to our findings, a Finnish study found that infants who had low weight gain in the first 6 months had higher risk for development of glucose intolerance, an effect that was greater for those with low birth weight [27]. Since our cohort excluded infants with birth weights below 3 kg, it is clear that the association we find between infant weight gain and adolescent MetS risk is independent of low birth weight. Furthermore, this association did not depend on family history of conditions related to the MetS such as type 2 diabetes, dyslipidemia, or myocardial infarction.
There is accumulating evidence that breastfeeding offers some protection related to the development of obesity, and that the effect may be “dose-dependent” [28, 29]. Because breastfed infants gain weight more slowly over the first year compared to formula-fed infants [30, 31], infant weight gain may pertain to the mechanism that decreases obesity risk in those who were breastfed. Having been breastfed has also been associated with lower risk for hypercholesterolemia, hypertension, diabetes, glucose intolerance, and insulin resistance [912]. To our knowledge, no other study has shown an association between breastfeeding and MetS risk in adolescence. Importantly, the significant effects of weight gain and breastfeeding were independent, suggesting that the effect of breastfeeding on MetS risk was not mediated by early infancy weight gain.
We do not know why males had higher MetS risk scores compared to females, but they had marginally significant higher birth weights and gained more weight in the first 3 months. Nonetheless, the effect of gender on MetS was independent of birth weight and infancy weight gain. This finding is consistent with higher prevalence rates of MetS in men compared to women in Chile [32]. In US adolescents, males are also more likely to have clustering of metabolic syndrome risk factors compared to girls [2]. However, in the Scandinavian study of infant weight gain and the MetS, male gender was not related to higher MetS risk [19], even though boys were similarly heavier at birth and gained more in infancy than girls. This suggests that the effect of gender is related to context rather than biology.
Our study has several limitations. The cohort was enrolled from a low- to middle-income community in Santiago, Chile, during a period of economic and nutritional transition. The setting and the fact that children with birth weights under 3 kg were not included limits generalizability. The study also has many strengths. The context of economic growth, high rates of breastfeeding, and nutritional support for infants allowed us to assess a sample where malnutrition was not a confounding factor. The longitudinal study took place at a nutrition research center allowing for detailed anthropometric measurement during infancy and the adolescent wave of data collection. Other strengths of the study include prospective data collection including monthly anthropometry in infancy and breastfeeding data collected from 4 to 12 months. In addition, the adolescent data collection included family history of diabetes, hypertension, elevated cholesterol, and heart attack.
5. Conclusion
In conclusion, this study adds to the current knowledge about early infant growth and breastfeeding and their long-term health effects. Higher infant weight gain was associated with increased MetS risk, whereas longer duration of exclusive breastfeeding was protective in healthy adolescents living in a rapidly developing country. Considering the increasing prevalence of the MetS in younger age groups and associations between the MetS and later disease, the replication and validation of these findings in different contexts is warranted.
Acknowledgments
The authors would like to express their gratitude to the participants and their families for their ongoing participation. The project was supported by grants from the National Heart, Lung, and Blood Institute (R01HL088530, PI: S. Gahagan) and the National Institute of Child Health & Human Development (R01HD14122 and R01HD33487, PI: B. Lozoff). The content of this paper is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Kozoru; Searching Only Hubs and Authorities
Sep 28, 2004 • 1:12 pm | (0) by | Filed Under Other Search Engines
All this talk in the forums on hubs and authorities, but yet no one really has built a search engine that searches exclusively the hubs and authorities. Of course, limiting the index to just include hubs and authorities would make for a less broad search engine. However, Kozoru, is looking to build a search service that searches hubs and authorities. Below you will find a quote being pulled from Brett Tabke at WebmasterWorld.
"We're going for a niche," said Kozuru Chief Executive John Flowers." We're specifically focusing on providing answers to specifically tailored questions.
"If you're asking a question about what car in 2003 had the highest safety rating, or how do I change the oil in my Acura, you can get those results in a regular search engine. But you'll have to sift through a lot of answers."
"We are trying to determine who the most authoritative sources are," he said.
Previous story: Ads On This Site
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Google Secures Webmaster Penalty Notifications within Webmaster Central
Jul 19, 2007 • 7:31 am | (1) by | Filed Under Google Search Engine Optimization
Google has released a new way for them to notify webmasters of possible penalties with their web sites.
In the past, Google sent emails, but this was abused by German hackers a couple times, so Google had to change how they handled that.
Now, if you are in violation of Google's web search TOS, you will receive a notification via Google Webmaster Tools, just one more reason to sign up and validate your site with Google Webmaster Central. If you are in violation, you will see a message indicator in the message center. It looks like this:
And if you click on a specific message, it will probably include content about search quality issues with a specific site. Google said, over time, they will also include non-search quality issue type of emails. Here is an example of a quality issue message:
If you login to your account and do not see a message, it does not mean you do not have quality search issues, at least today. Google said, "the number of sites we’re contacting is small, but we hope to expand this program over time." So, over time, hopefully all verified sites with quality issues will show a message in the message center.
Danny Sullivan explains that Google still may send out email notifications, but that will stop shortly. There is a current issue where if you get an email notification, and are not verified within Google Webmaster Central - you won't be able to see the message even after verifying your site. But Danny said they will be fixing that issue shortly.
Forum discussion at DigitalPoint Forums and WebmasterWorld.
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Daily Search Forum Recap: April 9, 10 & 13, 2009
Apr 13, 2009 • 4:00 pm | (0) by | Filed Under Search Forum Recap
Here is a recap of what happened in the search forums today, through the eyes of the Search Engine Roundtable and other search forums on the web.
Search Engine Roundtable Stories:
• Case Study Shows Pushed SEO Works Better Than Natural Site Development
A very helpful WebmasterWorld thread shares insights from a long time SEO who decided to pin two sites, competing in the same industry, with each other. He took two different strategies for each site. One site was pushed using old school SEO tactics, including doorway pages, paid and reciprocal links, content development and so on. The other site was to build a site with no paid links, unique content, no doorway pages, but add videos,
• Google Releases Maps for Mobile Update to Resolve Bugs
Google has announced a Google Maps for Mobile update in the Google Mobile Help discussions area that fixes the bugs with version 3.0 for Symbian and Windows Mobile devices. Googler, Christopher, said: A new version of Maps for mobile is available to download. This update includes many fixes and updates for both Maps and Google Latitude based on your feedback here in the Help Forum. Please see the following Release Notes for more details about
• Yahoo's New Local Targeting Feature Showing Poor Quality Traffic?
A month ago, Yahoo made some serious enhancements to the search marketing platform, improving the local targeting capabilities amongst other features. WebmasterWorld moderator, werty, has been gathering local data since the feature has been released and has posted the somewhat disappointing results. werty said in a WebmasterWorld thread: Currently we are using "North American YSM" and should only be getting North American traffic, but if I look through our logs I can see that 29%
• Google Still Showing Different Results Based on Query Case
Google continues to say that Google searches are "always case insensitive. Searching for [ new york times ] is the same as searching for [ New York Times ]." But since February 2008, we have been reporting cases where Google is case sensitive in terms of the search results they return. In fact, we reported it in February 2008, May 2008 and October 2008. So I am not surprised to see more threads about this
• Google AdSense Outage Resolved : April 2009
Late last week, Google AdSense had an issue where publishers were unable to login to the site and manage their ads. The reports began coming in on April 10th and some earlier reports came in on the night of the 9th. We have threads at Google AdSense Help, Google AdWords Help and two at WebmasterWorld and dozens at DigitalPoint Forums. Both AdSensePro Jennifer and AdWordsPro replied to the issue on the 10th. AdSensePro Jennifer chimed
• (Text) Recap of Weekly Search Buzz :: April 12, 2009
Due to the Passover holiday, I am only posting a text recap for the past week's coverage here at the Search Engine Roundtable. Google is making local queries more generic. There was link building prior to Google, see my write up on it, with Eric Ward's excellent comment. AdWords employee may have stepped over the line, ShoeMoney sues. Google testing a Twitter ad unit? Google ups the competitive ad filter. Publishers are still optimistic about
• Easter Logos From Search Engines But Not Google or Yahoo
Today is the holiday of Easter, it is also Passover. Some search engines have special logos and themes for the day, while others don't. Google, Yahoo & AOL are all missing special logos for Easter or Passover. Microsoft's Live.com, Ask.com, DogPile and a few others do have special Easter themes and logos. Here they are: Microsoft's Live.com theme: Ask.com: DogPile: Cre8asite Forums: Bruce Clay: Happy Holidays! Forum discussion at Cre8asite Forums.
• Optimism Abounds For Some Despite Adsense Earnings Drop
Some webmasters on WebmasterWorld are keeping an optimistic view on Adsense despite disappointment in their earning this year. The issues with Adsense lately are no secret, ranking from reporting issues to a Google glitch holding Adsense checks for some publishers. Let conspiracy theories on why abound, but that's not going to halt some from keeping a positive look on what an earnings drop in Adsense can actually spur you to do. For some when issues
• Does Google Look At Keywords In Long Titles?
Google typically cuts down the display of the title tag in the search results to no more than 65 characters. A WebmasterWorld thread asks does that mean Google won't look beyond the 65th character to determine what that page is about? Or if they do look beyond that 65th character, does Google deem the value of the words after the 65th character as less valuable? The title tag is arguably the most important on page
• Advertisers Want Google AdWords Time Zone Settings
A couple weeks ago, we polled our audience asking if Google should add time zone preferences to their AdWords feature list. Yahoo recently launched day parting, but gave advertisers the option to day part based on either the advertiser's time zone or the searcher's time zone. Google AdWords time zone setting is only based on your account's time zone, i.e. the advertiser's time zone. So if you have some one looking for your services and
• Google's Ranking My Images But From A Stolen Image Source
A WebmasterWorld thread has discussion started from a webmaster of a five year old e-commerce site, which had seen a drop in their Google Image search traffic, only to notice that their images are still in the Google results, but Google is using a different source to display the images. What that means is that Google thinks the other source is the true source of the images, while in fact, that source has stolen the
• Do SEOs Need To Know Coding To Be SEOs?
A heated Sphinn thread started some controversy in the SEO industry over who is an SEO and who is not an SEO. In short, Edward Lewis wrote a piece named HTML 4 SEO Best Practices for HTML Authoring. The article goes through many HTML attributes that can come in handy when coding your pages. The article is a very useful resource to hold on to and bookmark, if you need to look up the various
• Google Analytics Users Want Longer Data Storage For Free
Technically, Google has agreed to store your Google Analytics data for at least 25 months. We ran a poll the other week, asking you if that is long enough. Most of you said, no it is not. Of the 190 plus responses, 156 of your, or 82% said, that 25 months of storage is not enough time. While 35 of you, or 18% said it is enough. Truth be told, Google seems to store this
• Link Building Pre-Google Days
A WebmasterWorld thread has discussion on the topic of link building. This link building topic is different then the average thread. In this thread, the member asks if there was link building before Google became popular. The link building market, which is a niche within the niche of SEO, is almost completely driven by the popularity of the Google algorithm - at least these days. Typically, the more, high quality, relevant links you have to
• SEOs Split On If Store Discounts Are Equal to Link Buying
A few weeks ago, we wrote a story on how e-commerce sites can build links through offering customers a discount on orders, if they link back to the site. It is a neat way to get links but we wanted to poll our audience asking if they felt it was considered link buying. Since I am technically offline today, I thought it would be a good day to post the results of the survey. The
• SEO Doesn't Have To Look Bad
There is a big misconception out there that to make a site rank well in search engines, you need to make your site look ugly. SEO (search engine optimization) does not have to look ugly. Yes, having a lot of text and hyperlinking your content, where it makes sense, is important SEO factors, but it doesn't have to look ugly. A Google Webmasters Help thread has a webmaster who said that the SEO process is
• Reminder: Reverify Google Webmaster Tools Site Owners
Back in April 2007, Google gave webmasters a new easy way to remove content in Google, through Google Webmaster Tools. It is a great feature to quickly remove your content in Google, but it is an extremely powerful tool. If the wrong people gain access to your webmaster tools account, either by getting your username and password or by hacking into your site and validating your site on their webmaster tools account - you can
• Poll Result: How Long Does It Take To Rank Well?
A week ago, we polled our audience asking How Long Should It Take To Rank Well in Google? With over two-hundred responses, I thought it would be a good time to share the results with you all. How Long Does it Take To Rank Well in Google Results: :: 3 - 6 Months said 83 respondents or 35% :: 6 - 9 Months said 46 respondents or 19% :: 2 Months said 28 respondents or
Other Great Search Forum Threads:
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Place:Berkshire, Massachusetts, United States
NameBerkshire
Alt namesBerkshiresource: Getty Vocabulary Program
TypeCounty
Coordinates42.333°N 73.25°W
Located inMassachusetts, United States (1760 - )
Contained Places
Unknown
Loudon
Cemetery
Clarksburg Cemetery
Main Street Cemetery
Quaker Meetinghouse Grounds
Inhabited place
Adams Junction
Adams
Alander
Alford
Algerie Four Corners
Arnoldsville
Ashley Falls
Beachwood
Becket Center
Becket
Berkshire
Bonny Rigg Corners
Bowens Corners
Braytonville
Brier
Briggsville
Camp Ashmere
Camp Becket
Camp Chimney Corners
Camp Danbee
Camp Emerson
Camp Glenmere
Camp Greylock
Camp Jayson
Camp Kingsmont
Camp Lenore
Camp Mah-Kee-Nac
Camp Marion White
Camp Meadowlark
Camp Merrill
Camp Mohawk
Camp Muriel Flagg
Camp Romaca
Camp Russell
Camp Stevenson
Camp Sumner
Camp Taconic
Camp Witawentin
Camp Wyoma
Carey Corner
Cheshire Harbor
Cheshire
Clarksburg
Clayton
Cold Spring
Crane Lake Camp
Dalton ( 1700 - )
Drury
East Otis
East Sheffield
East Windsor
Eastern Summit
Eastover
Egremont Plain
Egremont
Farnams
Florida
Glendale
Great Barrington
Hancock
Hartsville
Hemlock Brook
Hephzibah Heights
Hinsdale
Hoosac Tunnel
Houghtonville
Housatonic
Interlaken
Konkapot
Lanesborough
Larrywaug
Lee
Lenox Dale
Lenox Station
Lenox
Line
Little Egypt
Mahkeenac Heights
Maple Grove
Mill River
Monterey
Montville
Mount Washington
New Ashford
New Boston
New Lenox
New Marlborough
North Adams
North Egremont
North Hancock
North Otis
Otis
Peru
Pittsfield ( 1740 - )
Richmond Furnace
Richmond
Risingdale
Rockdale Mills
Roosterville
Rosemont
Sandisfield
Savoy Center
Savoy
Shaker Village
Sheffield
South Lee
South Sandisfield
South Williamstown
Southfield
Stearnsville
Stevens Corner
Stockbridge
Sweets Corner
Tanglewood
Town Crest Village
Tyringham
Van Deusenville
Washington
West Becket
West New Boston
West Otis
West Pittsfield
West Stockbridge Center
West Stockbridge
West Summit
Whitcomb Summit
Williamstown ( 1750 - )
Williamsville
Windsor
Unknown
Blackinton
Greylock
South Egremont
source: Getty Thesaurus of Geographic Names
source: Family History Library Catalog
the text in this section is copied from an article in Wikipedia
Berkshire County is a non-governmental county located on the western edge of the U.S. state of Massachusetts. As of the 2010 census, the population was 131,219. Its largest city and traditional county seat is Pittsfield. The Berkshire Hills are centered on Berkshire County, and the county itself is often referred to simply as "the Berkshires".
Contents
Timeline
Date Event Source
1760 County formed Source:Red Book: American State, County, and Town Sources
1761 Court records recorded Source:Red Book: American State, County, and Town Sources
1761 Land records recorded Source:Red Book: American State, County, and Town Sources
1761 Probate records recorded Source:Red Book: American State, County, and Town Sources
1790 First census Source:Population of States and Counties of the United States: 1790-1990
1830 No significant boundary changes after this year Source:Population of States and Counties of the United States: 1790-1990
Population History
source: Source:Population of States and Counties of the United States: 1790-1990
Census Year Population
1790 30,291
1800 33,885
1810 35,907
1820 35,720
1830 37,835
1840 41,745
1850 49,591
1860 55,120
1870 64,827
1880 69,032
1890 81,108
1900 95,667
1910 105,259
1920 113,033
1930 120,700
1940 122,273
1950 132,966
1960 142,135
1970 149,402
1980 145,110
1990 139,352
Research Tips
External links
• Outstanding guide to Berkshire County, Massachusetts family history and genealogy (FamilySearch Research Wiki). History, towns and cities, county histories, birth, marriage, and death records, deeds, wills, court records, divorce records, naturalizations, tax records, warnings out, maps, and societies.
This page uses content from the English Wikipedia. The original content was at Berkshire County, Massachusetts. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Place:Lakeville, St. Joseph, Indiana, United States
Watchers
NameLakeville
TypeTown
Coordinates41.524°N 86.273°W
Located inSt. Joseph, Indiana, United States
source: Getty Thesaurus of Geographic Names
source: Family History Library Catalog
the text in this section is copied from an article in Wikipedia
Lakeville is a town south of South Bend in Union Township, St. Joseph County, Indiana, United States. The population was 786 at the 2010 Census. It is part of the South BendMishawaka, IN-MI, Metropolitan Statistical Area.
Research Tips
This page uses content from the English Wikipedia. The original content was at Lakeville, Indiana. The list of authors can be seen in the page history. As with WeRelate, the content of Wikipedia is available under the Creative Commons Attribution/Share-Alike License.
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Eclipse and XDoclet Tutorial
Earlier I mentioned an Eclipse plug-in for jBOSS that gives IDE functionality to jBOSS programming. An new article from DevX tells how to use XDoclet and Eclipse. XDoclet is a code generation engine that uses metadata within your code files to generate the uninteresting portions. For example, I've had my 462 class (large-scale distributed programming) use XDoclet with EJBs. You basically write the bean file and XDoclet generates all the rest of the EJB files from that, provided the proper metadata is included. This latest article runs through a EJB tutorial to show how Eclipse and XDoclet work together to generate code for servlet-driven EJB-based application.
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Australian Bureau of Statistics
Celebrating the International Year of Statistics 2013
ABS Home > Statistics > By Release Date
6202.0.40.001 - Labour Force, Teenage Employment and Unemployment, Australia, Preliminary - Data Report, Apr 2000
Previous ISSUE Released at 11:30 AM (CANBERRA TIME) 11/05/2000
Page tools: Print Page Print All RSS Search this Product
• About this Release
ABOUT THIS RELEASE
Contains unadjusted estimates of the labour force status of the civilian population aged 15-19, by attendance at school or a tertiary educational institution, for all States, Territories and Australia. It is produced monthly and is released at the same time as Labour Force, Australia, Preliminary(6202.0).
See also 6202.0.
© Commonwealth of Australia 2013
Unless otherwise noted, content on this website is licensed under a Creative Commons Attribution 2.5 Australia Licence together with any terms, conditions and exclusions as set out in the website Copyright notice. For permission to do anything beyond the scope of this licence and copyright terms contact us.
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Netizen Report: Halal-net Edition
This post also available in:
Español · Netizen Report: Edición Halal-net
Image via Flickr user ix4svs (CC BY-SA 2.0)
Most of this report was researched, written, and edited by Tom Risen, Weiping Li, Renata Avila, Rayna St, Sarah Myers and Rebecca MacKinnon.
After years of planning Iran appears to have laid the foundations for a national Internet network, separate from the global Internet, according to the U.S.-based researcher Collin Anderson who is preparing to release a report on his findings in the near future. Anderson has previously unearthed evidence of Iranian government plans to build what some officials have previously described as a “halal” or “clean” Internet. Based on news reports about Anderson's research, groups such as the U.S.-based Human Rights First warned last week that Iran may have taken “one more step towards fragmented access to information.” According to Anderson, the national service relies on components sold by China-based telecommunications company Huawei.
Some are speculating about the nature of the network: whether it will be completely disconnected from the global Internet, or whether it will instead emulate China’s Great Firewall which blocks access to many outside websites while enabling Chinese Internet users to communicate with the rest of the world. Your Middle East quoted Mohammad Soleimani, former Minister of Communication and Information Technology and now head of a parliamentary communication committee, who said the new network “will not cut access to the Internet…because it would amount to imposing sanctions on ourselves, which would not be logical. However, filtering will remain in place.”
Reporters Without Borders reports that government offices and civil service departments throughout the country were supposed to be connected to the new national network on September 22. The actual launch of the network is still pending. Meanwhile, Iran has banned Gmail and to a lesser degree Google search, which officials said was to protest the anti-Islam film Innocence of Muslims. Earlier this month Iran launched a national email service, Iran.ir, which requires extensive information about the user's identity to join.
Censorship
The controversial film, Innocence of Muslims, continues to be used as an excuse by governments to block access to YouTube. Among the countries that have blocked YouTube pages are Bangladesh, Kyrgyzstan, and Afghanistan. Some Chechen Internet service providers have also blocked access to YouTube, but a court will now determine whether the move was legal. The case is testing the implementation of a new law in Russia to censor websites the government finds disagreeable or indecent. Communications Minister Nikolai Nikiforov has threatened to shut down YouTube nationwide using the law.
Meanwhile, in Los Angeles, Superior Court Judge Luis Lanvin denied a request to remove the controversial video from YouTube. The request was made by Cindy Lee Garcia, who acted in the film and claimed she was misinformed and misled about its actual content. YouTube itself blocked access to the film in Egypt, Libya, India and elsewhere.
As Myanmar loosens censorship, government officials and journalists have started talking about the way forward. One journalist from International Herald Tribune had the chance to meet his former censor and speak about the censorship process in Myanmar.
Jordan’s King Abdullah endorsed a new media law which requires “electronic publications” to be licensed by the government. Critics say it could restrict freedom of expression.
The US Patent and Trademark Office (USPTO) blocked some websites within its offices, including those of the Electronic Frontier Foundation, Center for Democracy and Technology, and American Civil Liberties Union because the locations were dubbed “political” or “activist,” but then stopped filtering after the news spread and provoked criticism.
Thuggery
Syrian citizen journalist Abdel Karim al-Oqda, who used the pseudonym Abu Hassan to report from Hama, Syria, was burned to death after regime forces targeted his home.
Famous United Arab Emirates (UAE) blogger and political activist Ahmed Mansoor was beaten by an unidentified man at the university where he studies law.
Three founding members of Vietnam’s Free Journalists’ Club, a group of citizen journalists who advocate free speech and independent journalism, have been sentenced to jail for distributing “anti-state propaganda.”
In China, the famous Internet writer and former journalism professor Jiao Guobiao has been detained by Beijing’s public security bureau for “inciting subversion of state power” after he published articles on the Diaoyu Islands (known as “Senkaku” in Japan) territory dispute between China, Taiwan and Japan.
Surveillance
After recent violence in Assam, the Indian government is planning to set up a dedicated surveillance agency to monitor the Internet for false rumors and malicious content and warn national security agencies beforehand so that preventive measures can be adopted.
German Foreign Minister Guido Westerwelle spoke publicly against the sale of spying technology to repressive regimes. Several European political figures such as French Secretary of State for the Digital Economy, members of the Dutch Green Party and the EU Commissioner for the Digital Agenda have also expressed their concern over European technology corporations selling spyware to authoritarian countries.
In New Zealand, the Prime Minister John Key has asked the Inspector-General of Intelligence and Security to investigate a case in which New Zealand’s intelligence agency helped the US government to illegally spy on Megaupload founder Kim Dotcom and other people involved in this case.
Privacy
The US Federal Trade Commission (FTC) has proposed changes to the Children’s Online Privacy Protection Act (COPPA) which could raise legal risks to plugin program developers and website operators. According to the proposals, plugin developers could face legal liability for collecting childrens’ information if they receive IP addresses from plugins installed on childrens’ websites; another proposal seeks to expand the definition of sites “directed to children.”
The latest Facebook audit review by the Irish Data Protection Commissioner shows that Facebook has decided to turn off its controversial facial recognition feature and related data in Europe on October 15. The decision applies to the “tag suggestion” feature which automatically tag names on faces in the pictures when users upload photos to Facebook.
The browser feature “Do Not Track”, which prevents advertisers from tracking users’ online activities, has been added to several browsers such as Firefox, Internet Explorer, Apple Safari, Opera. Google has also promised to add this function by the end of this year. Here is a guide on how to enable “Do Not track” in various browsers.
The “cryptoparty” concept has spread from Australian cities such as Melbourne to international locations such as London, as people aware of privacy issues are getting together to learn about anonymity techniques and how to protect their right to privacy.
Cybersecurity
According to a recently unsealed court document in a computer fraud case that Microsoft has filed against Chinese Web domain 3322.org, some new computers had malware installed and were ready to attack websites from the moment they’re turned on. The malware may come from counterfeit software which some Chinese computer manufacturers had used to save costs.
To cope with the security breach dubbed “CRIME” (Compression Ratio Info-leak Made Easy), which targets HTTP sessions, researchers have suggested that website operators should turn off a bandwidth-saving compression feature.
Security researcher Chris Soghoian recently discovered that a company called Packet Forensics is developing equipment that could intercept secure communications by using forged website security certificates without breaking encryption, and then marketing the equipment to law enforcement agencies. The boxes would have to be connected to an Internet service provider by a law enforcement agency and then persuade a Certificate Authority such as Verisign to collaborate.
National Policy
The Director of Public Prosecutions in the UK plans to issue guidelines to help prosecutors decide whether to press charges in cases involving social media. The agency has announced they will start public consultation procedures.
A leaked document of the European Commission-funded “Clean IT” project, which aims to combat online terrorist content by encouraging cooperation between governments and Internet companies, has revealed that EU officials have proposed extreme measures such as authorizing the police to “patrol” the Internet and strengthening regulations to prevent anonymous use of online services.
Copyright
According to the 2012 International Piracy Watch List released by the US Congressional International Anti-Piracy Caucus (IAPC), Switzerland and Italy are now on the list of countries of concern along with countries such as Russia, China, and Ukraine. IAPC said Switzerland’s inadequate copyright law has made it “a home for rogue sites,” and that Italy needs substantial copyright reforms to combat illegal downloading.
Sovereigns of Cyberspace
A trustee of the Wikimedia Foundation UK and a high-level editor at Wikipedia have been discovered to have edited Wikipedia’s “Did You Know” feature and other projects to benefit their private clients for more exposure on Wikipedia pages. The trustee has resigned after the revelation of the pay-to-play relationship.
Recently Google was granted a US patent for a technology that allows users to create multiple pseudonyms online and decide when to reveal true identities to others. However experts have held different positions toward this development: some have worried Google could gather more sensitive personal information, while others have thought it is positive to have different levels of online identities for online activities.
Google has shut down its free online music download service in China in order to put more resources on “high impact products.” Google’s music service is one of the few remaining services on its China-based landing page after the company moved its search engine servers to Hong Kong in 2010.
Internet companies including Google, Facebook, AOL, Yahoo! and LinkedIn have formed an organization named “The Internet Association” to coordinate their efforts in lobbying for Internet freedom and economic issues in Washington D.C.
Internet Governance
Freedom House has published the report “Freedom on the Net 2012: A Global Assessment of Internet and Digital Media.” The report argues that governments around the world have grown more sophisticated in controlling the flow of online information. At the same time, civil societies have fought back and won several important victories. The report says that people in Estonia enjoy the greatest degree of Internet freedom, while people in Iran, Cuba and China have the lowest among the countries examined.
The International Telecommunication Union (ITU), an agency of the United Nations, released the “State of Broadband 2012” report on international Internet usage data. It showed that currently one-third of world's population has access to the Internet. The nation with the highest Internet usage rate is Iceland.
Cool Things
Ecuador will become a “free software” [es] territory, President Rafael Correa recently announced.
Riot police had to be called to the town of Haren in The Netherlands after a teenager’s Facebook event reportedly attracted 30,000 people to the tiny town of 18,000 inhabitants.
Publications and Studies
Subscribe to the Netizen Report by email
For upcoming events related to the future of citizen rights in the digital age, see the Global Voices Events Calendar.
Join the conversation
Authors, please log in »
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293 reputation
14
bio website ozzysoft.com
location
age
visits member for 1 year, 8 months
seen 2 days ago
stats profile views 16
I've been a professional developer since 1992, and started as a hobbyist back in 1982. As a hobbyist in the 80's and early 90's I worked with TRS80, Apollo, Dec, Apple II, Mac, PC's. Starting as a professional in the early 90's, I've worked with OS/2, Windows, and Linux, both on the web, client-server and multi-tier business applications, working mainly with Microsoft technologies through my career. I've been dabbling with Android lately (which has rekindled my hobbyist spirit!) I'm currently employed working on investment banking analytics.
Released my first android app! check it out: http://www.mazerace.com
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101 reputation
2
bio website twitter.com/ricmarques
location Portugal
age 36
visits member for 8 months
seen Sep 4 '12 at 13:50
stats profile views 0
Nowadays, professionally I'm mainly a Linux server admin (mostly SUSE - openSUSE and SLES) of Testing, Development and Production servers (both physical servers and virtual machines - usually VMware). So, I work regularly with VMware products, namely vSphere (ESXi 4.1 and vCenter Server 4.1).
My netbook runs Ubuntu and sometimes I "play" with Debian systems.
The coding that I do is mostly in Perl and "shell scripts" (for the "bash" shell) for automating system administration tasks in Linux servers. In "previous lives", I coded web applications in PHP for LAMP based projects and also in ASP ("Active Server Pages"), both "classic ASP" - ASP + VBScript - and ASP.Net - using C#
Certications:
- RHCE - "Red Hat Certified Engineer"
- VCP - "VMware Certified Professional" ("VCP4" and "VCP5")
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Vol 5, No 2 (2012)
Vol. 5, No. 2, February 2012
Table of Contents
Articles
Teaching Writing Skills Based on a Genre Approach to L2 Primary School Students: An Action Research PDF
Hyejeong Ahn p2
Self-esteem and Academic Success as Influenced by Reading Strategies PDF
Eghbal Zarei, Nasrin Shokrpour, Elham Nasiri, Reza Kafipour p17
Empirical Study of Formative Evaluation in Adult ESL Teaching PDF
Qin Yi p27
Formulaic Sequences and Their Relationship with Speaking and Listening Abilities PDF
Ebrahim Khodadady, Saeedeh Shamsaee p39
Non-native Student’s Communication is Affected Due to the Lack of Pragmatic Competence PDF
Latha V. G., Premalatha Rajan p50
An Exploration of Private Language Institute Teachers’ Perceptions of Written Grammar Feedback in EFL Classes PDF
Mina Jodaie, Farahman Farrokhi p58
Are Modal Auxiliaries in Malaysian English Language Textbooks in Line with Their Usage in Real Language? PDF
Laleh Khojasteh, Reza Kafipour p68
The Pragmatic Knowledge of Iranian EFL Learners in Using Face Keeping Strategies in Reaction to Complaints at Two Different Levels PDF
Akram Azarmi, Biook Behnam p78
Apology Strategies of Iranian Undergraduate Students PDF
Mohammad Dadkhah Tehrani, Omid Rezaei, Salman Dezhara, Reza Soltani Kafrani p93
A Survey on the Iranian ELT Community's Attitudes to Critical Pedagogy PDF
Hossein Davari, Abutaleb Iranmehr, Seyyed Mahdi Erfani p101
ESP Practitioner Professionalization through Apprenticeship of Practice: the Case of Two Iranian ESP Practitioners PDF
Batoul Ghanbari, Abbas Eslami Rasekh p112
On the Effect of Gender and Years of Instruction on Iranian EFL Learners' Collocational Competence PDF
Mansoor Ganji p123
Gender Bias in the Iranian High School EFL Textbooks PDF
Mohadeseh Amini, Parviz Birjandi p134
Binding Task-Based Language Teaching and Task-Based Language Testing: A Survey into EFL Teachers and Learners' Views of Task-Based Approach PDF
Ali Panahi p148
Vocabulary Learning Strategies of Medical Students at Shiraz University of Medical Sciences PDF
Fatemeh Seddigh, Nasrin Shokrpur p160
The Relationship between Iranian EFL Learners’ Self-efficacy Beliefs and Use of Vocabulary Learning Strategies PDF
Farrokhlagha Heidari, Mehri Izadi, Mansooreh Vahed Ahmadian p174
This work is licensed under a Creative Commons Attribution 3.0 License.
English Language Teaching ISSN 1916-4742 (Print) ISSN 1916-4750 (Online)
Copyright © Canadian Center of Science and Education
To make sure that you can receive messages from us, please add the 'ccsenet.org' domain to your e-mail 'safe list'. If you do not receive e-mail in your 'inbox', check your 'bulk mail' or 'junk mail' folders.
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"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://ccsenet.org/journal/index.php/ijbm/article/view/8077"
}
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On Key Points of Financial Cost Management in Small and Medium-Sized Enterprises
Xingwu Yu, Weixing Wang
Abstract
Chinese small and medium-sized enterprises (SME), accounting for 99.3% of all Chinese corporations, create
enormous economic fortune and employment opportunity. However, most of SMEs are lack of capital, with
simple equipments and improper management, so they are incapable to withstand risks and easy to be involved
in difficulty. Therefore, we should properly guide SMEs to enhance financing, investment management and the
management of other value chain activities, which will contribute to their survival, development, strength, and
increasing improvement of market competitiveness.
Full Text: PDF
This work is licensed under a Creative Commons Attribution 3.0 License.
International Journal of Business and Management ISSN 1833-3850 (Print) ISSN 1833-8119 (Online)
Copyright © Canadian Center of Science and Education
To make sure that you can receive messages from us, please add the 'ccsenet.org' domain to your e-mail 'safe list'. If you do not receive e-mail in your 'inbox', check your 'bulk mail' or 'junk mail' folders.
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Connexions
Sections
You are here: Home » Content » Preparing a Word Document for Creating a Connexions Module
About: Preparing a Word Document for Creating a Connexions Module
Module by: Kenneth Leroy Busbee. E-mail the author
View the content: Preparing a Word Document for Creating a Connexions Module
Metadata
Name: Preparing a Word Document for Creating a Connexions Module
ID: m33255
Language: English (en)
Summary: Detailed help on how to prepare a Word document for importing into a Connexions module. Use of both Word styles and commonly used Connexions' CNXML styles are demonstrated.
Subject: Science and Technology
License: Creative Commons Attribution License CC-BY 3.0
Authors: Kenneth Leroy Busbee (ken.busbee@hccs.edu)
Copyright Holders: Kenneth Leroy Busbee (ken.busbee@hccs.edu)
Maintainers: Kenneth Leroy Busbee (ken.busbee@hccs.edu)
Latest version: 1.3 (history)
First publication date: Dec 22, 2009 2:27 pm -0600
Last revision to module: Jan 13, 2010 8:12 pm -0600
Downloads
PDF: m33255_1.3.pdf PDF file, for viewing content offline and printing. Learn more.
EPUB: m33255_1.3.epub Electronic publication file, for viewing in handheld devices. Learn more.
XML: m33255_1.3.cnxml XML that defines the structure and contents of the module, minus any included media files. Can be reimported in the editing interface. Learn more.
Source Export ZIP: m33255_1.3.zip ZIP containing the module XML plus any included media files. Can be reimported in the editing interface. Learn more.
Version History
Version: 1.3 Jan 13, 2010 8:12 pm -0600 by Kenneth Leroy Busbee
Changes:
Made several typo corrections.
Version: 1.2 Dec 22, 2009 3:57 pm -0600 by Kenneth Leroy Busbee
Changes:
Corrected downloading hyperlinks for the four files.
Version: 1.1 Dec 22, 2009 3:05 pm -0600 by Kenneth Leroy Busbee
Changes:
Initial module.
How to Reuse and Attribute This Content
If you derive a copy of this content using a Connexions account and publish your version, proper attribution of the original work will be automatically done for you.
If you reuse this work elsewhere, in order to comply with the attribution requirements of the license (CC-BY 3.0), you must include
• the authors' names: Kenneth Busbee
• the title of the work: Preparing a Word Document for Creating a Connexions Module
• the Connexions URL where the work can be found: http://cnx.org/content/m33255/1.3/
See the citation section below for examples you can copy.
How to Cite and Attribute This Content
The following citation styles comply with the attribution requirements for the license (CC-BY 3.0) of this work:
American Chemical Society (ACS) Style Guide:
Busbee, K. Preparing a Word Document for Creating a Connexions Module, Connexions Web site. http://cnx.org/content/m33255/1.3/, Jan 13, 2010.
American Medical Assocation (AMA) Manual of Style:
Busbee K. Preparing a Word Document for Creating a Connexions Module [Connexions Web site]. January 13, 2010. Available at: http://cnx.org/content/m33255/1.3/.
American Psychological Assocation (APA) Publication Manual:
Busbee, K. (2010, January 13). Preparing a Word Document for Creating a Connexions Module. Retrieved from the Connexions Web site: http://cnx.org/content/m33255/1.3/
Chicago Manual of Style (Bibliography):
Busbee, Kenneth. "Preparing a Word Document for Creating a Connexions Module." Connexions. January 13, 2010. http://cnx.org/content/m33255/1.3/.
Chicago Manual of Style (Note):
Kenneth Busbee, "Preparing a Word Document for Creating a Connexions Module," Connexions, January 13, 2010, http://cnx.org/content/m33255/1.3/.
Chicago Manual of Style (Reference, in Author-Date style):
Busbee, K. 2010. Preparing a Word Document for Creating a Connexions Module. Connexions, January 13, 2010. http://cnx.org/content/m33255/1.3/.
Modern Languages Association (MLA) Style Manual:
Busbee, Kenneth. Preparing a Word Document for Creating a Connexions Module. Connexions. 13 Jan. 2010 <http://cnx.org/content/m33255/1.3/>.
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"url": "dungeons.wikia.com/wiki/SRD:Dissipater?direction=prev&oldid=9351",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://dungeons.wikia.com/wiki/SRD:Dissipater?direction=prev&oldid=9351"
}
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Wikia
SRD:Dissipater
Talk0
9,503pages on
this wiki
Revision as of 02:33, April 23, 2009 by Dmilewski (Talk)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
This material is published under the OGL
Dissipater: This kind of weapon is devastating to creatures and objects composed of or originally formed from ectoplasm (such as astral constructs, walls of ectoplasm, creatures in ectoplasmic form, and items created using the metacreativity discipline). Against qualifying targets, a dissipater weapon ignores damage reduction and hardness, and treats all successful hits as critical hits.
Strong metacreativity; ML 12th; Craft Psionic Arms and Armor, dismiss ectoplasm; Price +1 bonus.
Back to Main PageSystem Reference DocumentPsionic Items
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"url": "elinux.org/index.php?oldid=27487&title=Didj_and_Explorer_libSDL",
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"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://elinux.org/index.php?title=Didj_and_Explorer_libSDL&oldid=27487"
}
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Didj and Explorer libSDL
From eLinux.org
Revision as of 02:37, 19 November 2010 by Julspower (Talk | contribs)
Jump to: navigation, search
libSDL is a popular library for games and other applications. It provides APIs for events (buttons, etc), graphics, sound, and more.
Note: A framebuffer driver is required. **not strictly true, will still run audio without a framebuffer**
Create an install script for iconv:
create a folder in your packages dir called libiconv, then copy and paste the following into a text file and save it in the libiconv folder, call it install.sh (you might need to change permissions on the file as well).
Download libiconv-1.13.1.tar.gz google it pretty easy to find
#!/bin/bash
BUILD_FROM_SOURCE=1
PKG_NAME=libiconv
SRC=libiconv-1.13.1.tar.gz
set -e
. $PROJECT_PATH/scripts/functions
# make sure all of the environment variables are good
check_vars
# exit if the user is root
check_user
# parse args
set_standard_opts $*
pushd $PROJECT_PATH/packages/$PKG_NAME
BUILD_DIR=`echo "$SRC" | cut -d '.' -f -3`
if [ "$CLEAN" == "1" -o ! -e $BUILD_DIR ]; then
rm -rf $BUILD_DIR
tar -xzf $SRC
fi
pushd $BUILD_DIR
CFLAGS="-I$PROJECT_PATH/packages/$PKG_NAME/libiconv-1.13.1/ -O3 -fPIC -mcpu=arm926ej-s" LDFLAGS=-L$PROJECT_PATH/packages/$PKG_NAME/libiconv-1.13.1/ ./configure --host=arm-linux --target=arm-linux --prefix=$ROOTFS_PATH/usr --enable-shared
make clean
make
make install
popd
popd
exit 0
Build libiconv
Now that you've got an install script you can run it with the following command:
CLEAN=1 ./install.sh
This will download the package for you, unpack it, set the necessary vars then compile and install libiconv. You should only need to do CLEAN=1 for the first time you run it, that's the var that triggers a clean download and install.
Build the SDL library
To build SDL, first download and unpack it:
wget http://www.libsdl.org/release/SDL-1.2.13.tar.gz
tar -xf SDL-1.2.13.tar.gz
cd SDL-1.2.13
You'll need a toolchain, for example this one
Now try the following, but replace the "/path/to" lines with paths on your system. You'll want to tell the build system where to install libSDL and where to look for kernel headers
CFLAGS="-I/path/to/kernel/linux-2.6/include/"
CC=arm-linux-uclibcgnueabi-gcc
CXX=arm-linux-uclibcgnueabi-g++
./configure --prefix=/path/to/rootfs//usr/ --build=`uname -m` --host=arm-linux --disable-video-opengl --disable-video-x11
--disable-esd --disable-video-directfb --enable-video-fbcon --enable-pulseaudio=no
If that succeeds, try building and installing:
make
make install
You should see libSDL.so in usr/lib in your rootfs directory as well as its header files in "usr/include". When building applications against libSDL, tell the compiler to look for headers and the linker to look for libraries there (ie: CFLAGS="-I/path/to/rootfs/usr/include" and LDFLAGS="-L/path/to/rootfs/usr/lib").
Set up the device
Copy the .so files and symlinks to your device. Also edit "/etc/profile" on your device and add some environment variables for SDL:
For Didj:
export SDL_NOMOUSE=1
export SDL_FBDEV="/dev/fb0"
export SDL_VIDEODRIVER="fbcon"
export SDL_AUDIODRIVER="dsp"
export SDL_PATH_DSP="/dev/dsp"
export SDL_DEBUG="1"
export SDL_FBACCEL="0"
For Explorer, the touchscreen works sort of like a mouse (TODO: add tslib support?), so you could try:
export SDL_MOUSEDEV="/dev/input/event3"
export SDL_FBDEV="/dev/fb0"
export SDL_VIDEODRIVER="fbcon"
export SDL_AUDIODRIVER="dsp"
export SDL_PATH_DSP="/dev/dsp"
export SDL_DEBUG="1"
export SDL_FBACCEL="0"
You'll need to source that on your device:
. /etc/profile
or just reboot. You should now be able to run SDL programs or demos as long as your device is set up with a standard Linux framebuffer and console in place of the LeapFrog-supplied graphics drivers.
TODO
* better audio support?
* tslib to make touchscreen usable on Explorer?
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{
"content_type": "text/html",
"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31772",
"uncompressed_offset": 101900174,
"url": "googlesystem.blogspot.com/2007/10/traffic-analysis-for-content-hosted-by.html",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://googlesystem.blogspot.com/2007/10/traffic-analysis-for-content-hosted-by.html"
}
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An unofficial blog that watches Google's attempts to move your operating system online.
Send your tips to gostips@gmail.com.
October 22, 2007
Traffic Analysis for Content Hosted by Google
Google Analytics is useful to find information about your site's traffic, but it would be even more useful to gather stats for other sites you use to post your content online. For example, you might post videos on YouTube, upload photos to Picasa Web Albums and create documents using Google Docs. It would be interesting to link these services with Google Analytics and get access to a wealth of information about the number of views for each item, the time spent on a page, the referrals and more.
Google already does this for Project Hosting: "To provide the most useful software to your users, you might want to know simply how many potential users have visited your project workspace, which countries they come from, which browsers they use, and which of your wiki pages they have viewed. Now all those questions can be answered. Project owners may simply sign up for Google Analytics and enter an analytics profile number into the project admin page. Tracking data can be viewed on the Google Analytics site about 24 hours later."
This could solve a lot of user requests and make these services more personal and more useful. Scribd, a document sharing service I mentioned earlier this year, shows a lot of traffic data publicly, but I'm not sure if that's the way to go.
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{
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"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31777",
"uncompressed_offset": 107726573,
"url": "hitchwiki.org/en/Bad_Bentheim",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://hitchwiki.org/en/Bad_Bentheim"
}
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Bad Bentheim
From Hitchwiki
Jump to: navigation, search
See full map
Bad Bentheim is a city in the southwest of Lower Saxony, close to the Dutch border.
Near this city the motorway A30 starts.
If you're in Bad Bentheim, don't hesitate to visit the castle, or at least have a look at it. It's nice.
Hitchhiking spot westwards, near the train station.
Hitchhiking out
West towards Gronau, Oldenzaal (Netherlands)
If you travel by a Regionalbahn (train) towards the Netherlands, you may have to wait several hours on this station for the Intercity towards Amsterdam. From the station you need to hike about 300 meters south to get on a crossing with a bay where you may easily hitchhike to Gronau, Oldenzaal or Hengelo (see image). Although it's only some kilometers towards Oldenzaal, there are quite few cars going that direction. You might wait a bit longer after passing Gildehaus. Towards Gronau, it shouldn't be such a big problem. This spot is much better than the next bus bay at the end of town, because it's behind the traffic lights, so traffic is not splitting up. Try to catch a ride towards Gildehaus at least, and don't get stuck at the next traffic light, as there are no good spots.
North towards Nordhorn, A30
At the crossing, just take the road towards the north or try it behind the train station. The street (403) directly goes towards the on ramp Nordhorn/Bad Bentheim.
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{
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"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31784",
"uncompressed_offset": 137925340,
"url": "lists.maemo.org/pipermail/maemo-developers/2008-December/018009.html",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://lists.maemo.org/pipermail/maemo-developers/2008-December/018009.html"
}
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[maemo-developers] g_enum_get_value_by_name
From: Pekka Pessi Pekka.Pessi at nokia.com
Date: Thu Dec 18 09:36:21 EET 2008
Arto Karppinen <arto.karppinen at mail.suomi.net> writes:
>I need to save and read an enum from a GKeyFile. I was going save the
>value as a string into the keyfile and use g_enum_get_value_by_name() to
>parse the value, but i cant figure out how to get the GEnumClass parameter.
>GEnumValue* g_enum_get_value_by_name(GEnumClass *enum_class,
> const gchar *name);
g_type_class_ref() and then unref or if you already have ref'ed,
g_type_class_peek().
--Pekka
More information about the maemo-developers mailing list
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{
"content_type": "text/html",
"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31809",
"uncompressed_offset": 171463332,
"url": "openwetware.org/index.php?redirect=no&title=Protein_Purification_-_Salting_Out",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://openwetware.org/index.php?title=Protein_Purification_-_Salting_Out&redirect=no"
}
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Protein Purification - Salting Out
From OpenWetWare
Jump to: navigation, search
Different proteins salt out at different concentrations of ammonium sulfate. Here are some references on salting out and tools for calculating the amounts needed.
Current Protocols in Protein Science, appendix 3F contains information on salting out. DOI:10.1002/0471140864.psa03fs13
Personal tools
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{
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"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31810",
"uncompressed_offset": 171490505,
"url": "openwetware.org/index.php?curid=123136&diff=0&oldid=632353&title=Sack%3AProtocol_Pdfs",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://openwetware.org/index.php?title=Sack:Protocol_Pdfs&curid=123136&diff=0&oldid=632353"
}
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Sack:Protocol Pdfs
From OpenWetWare
(Difference between revisions)
Jump to: navigation, search
Current revision (23:00, 3 October 2012) (view source)
Line 18: Line 18:
*To view any of these protocols on the wiki go to [[Sack:Protocols]]
*To view any of these protocols on the wiki go to [[Sack:Protocols]]
-
[http://openwetware.org/images/2/27/Cell_Media_with_Gen_Selection_Agents_4-5-2012.docx=0 download]
+
-
[[http://openwetware.org/images/2/27/Cell_Media_with_Gen_Selection_Agents_4-5-2012.docx download]]
+
[http://openwetware.org/images/2/27/Cell_Media_with_Gen_Selection_Agents_4-5-2012.docx download]
*Return to [[Sack]] Homepage
*Return to [[Sack]] Homepage
Current revision
Cell Culture
Cell Media with Selection Agents (Blasticidin, Zeocin, Geneticin) 04/05/12 Pdf
Cell Media with Selection Agents (Blasticidin, Zeocin) 11/15/11Pdf
Cell Splitting Protocol 03/02/12Pdf
Cell Transfection (Lipofectamine LTX) 05/01/12Pdf
Cell Transfection (Lipofectamine) 08/14/12Pdf
Cell Transfection (Novachoice) 05/04/12Pdf
Cell Harvesting for Electrophysiology 07/18/12Pdf
Cell Plating for Electrophysiology 10/21/11Pdf
Freezing CHO Cells 04/05/12Pdf
Plating Subclones for Ephys 09/29/11Pdf
Plating Subclones for Flourescent Screening 04/05/12Pdf
Subcloning Cells 02/23/11 Pdf
Testing TREX Subclones Ephys 04/06/11Pdf
Thawing CHO cells 07/09/12Pdf
download
• Return to Sack Homepage
Personal tools
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{
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"provenance": "cccc-CC-MAIN-2013-20-0000.json.gz:31812",
"uncompressed_offset": 171524073,
"url": "openwetware.org/wiki/User:Karlena_L._Brown/Notebook/PVOH_Research/2013/02/04",
"warc_date": "2013-11-22T19:23:19.000Z",
"warc_filename": "<urn:uuid:b736a179-817d-4f61-b1de-58c80ec9d549>",
"warc_url": "http://openwetware.org/wiki/User:Karlena_L._Brown/Notebook/PVOH_Research/2013/02/04"
}
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User:Karlena L. Brown/Notebook/PVOH Research/2013/02/04
From OpenWetWare
Jump to: navigation, search
PVOH Research Main project page
Previous entry Next entry
OBJECTIVES
ACCIDENTALLY PLACED THIS ENTRY ON THE FEBRUARY 4th; HOWEVER, THIS ENTRY SHOULD BE UNDER FEBRUARY 6th. THANK YOU.
• Begin preparing polyvinyl alcohol microsperes for experimental analysis
• Filter surfactant exchanged modified clays
• Complete freeze and thaw crosslinked clay cycle #1
90:10 PVOH Clay Microsphere Preparations
*Reaction should be carried out using a reflux condenser and a thermostatic water bath of 55°C.
1. Dissolve ~ 1.0g total of PVOH 146K and clay additive selected (laponite) in 5mL hot deionized H2O
2. Acidify the solution using 0.25mL 0.5M H2SO4
3. Dissolve ~ 0.6g cellulose acetate butyrate (CAB) in 25mL 1,2-dichloroethane within a round bottom flask
4. Pour the acidified PVOH 146K solution into the 25mL 1,2-dichloroethane solution within the clean round bottom flask
5. Stir 1,2-dichloroethane, CAB, and PVOH 146K solution for 30 minutes
6. After 30 minutes of stirring, add 0.75mL glutaraldehyde to the solution
7. After the addition of glutaraldehyde, allow the reaction to carry out for 3 additional hours at 55°C
8. Filter the produced microspheres from the solution using a a sintered glass filter and a vacuum
9. Wash the collected microspheres using ~ 2-3mL of the following solvents in the following order:
* 1,2-dichloroethane
* acetone
* hot water
* cold water
* methanol
1. Scrape off all of the collected microspheres into a small glass vial. Leave the lid off of the small glass vial.
2. Freeze microspheres in liquid nitrogen for ~ 2 minutes
3. Place on [ask Dr. Hartings name of machine again] overnight to completely dry
* The following procedure was adapted from the article Poly(vinyl alcohol) microspheres with pH- and thermosensitive properties as temperature-controlled drug delivery [1]
Preparation of Laponite Microspheres using PVOH MW 146,000-186,000 '
PVOH 146K added(g)0.9001
Laponite clay added (g)0.1004
Cellulose Acetate Butyrate added (g)0.6066
Glutaraldehyde added (mL)0.75
H2SO4 added (mL)0.25
Important 1,2 Dichloroethane and Glutaraldehyde Safety
• Glutaldehyde is toxic colorless liquid (MW: 100.12 g / mole)
• 1,2 Dichloroethane is a toxic colorless flammable liquid (MW: 98.96 g / mole)
• Avoid all glutaldehyde and 1,2 dichloroethane skin contact by wearing gloves and googles
• Glutaldehyde should be kept in the freezer when not in use
• 1,2 Dichloroethane should be kept in a flammable cabinet when not in use
• When in use, both glutaldehyde and 1,2 dichloroethane should be kept in the fume hood
• 1,2 Dichloroethane should be capped at all times when not in use to inhibit inhalation and free floating volatile particles in the air
• Keep the glutaldehyde vial in a beaker so that it does not spill
• Keep anything and everything containing some form of 1,2 dichloroethane in the hood and allow all residue to evaporate off before washing
• If spilled, wipe up glutaldehyde with paper towel while wearing gloves
• If spilled, use a spill kit towel to soak up excess 1,2 dichloroethane. Allow some of the residual essence of 1,2 dichloroethane to evaporate; then use acetone to wipe over area contaminated with 1,2 dichloroethane.
• Keep all spills of glutaldehyde and 1,2 dichloroethane contained in the fume hood
• Wash hands after removing gloves that were in contact with glutaldehyde or 1,2 dichloroethane
Clay Exchanged Solutions & Vacuum Filtration Procedures
The clays being filtered are the clays that were prepared on 2/1/13
Setup of a vacuum filtration system:
* Clean 250mL Erlenmeyer Filter Flask
* Large Buchner Funnel
* Whatman #42 Filter Paper
* Long Secure Rubber Hose
1. Filter a clay solution selected using the vacuum filtration system setup
2. Attach all parts of the apparatus to the vacuum filtrator system
3. Place a piece of Whatman #42 filter paper in the Buchner Funnel
4. Turn on the vacuum filtrator system
5. Check the suction rate to ensure that enough pressure is applied
6. Pour an exchanged clay solution into the Buchner funnel
7. Allow vacuum filtration to continue until the clay is completely dry
8. Discard the aqueous filtrate solution and place it in a labeled waste container
9. Collect the soil filtrate on the filter paper and scrape it into a small labeled metal pan
10. Place metal pan with the soil filtrate in the oven to dry for ~ 2-3 days
11. Wash all glassware and equipment first with H2O and then with acetone for later use
Notes
ACCIDENTALLY PLACED THIS ENTRY ON THE FEBRUARY 4th; HOWEVER, THIS ENTRY SHOULD BE UNDER FEBRUARY 6th. THANK YOU.
• Clays previously prepared on 2/1/13 only exchanged for ~ 5 days rather than a week.
• While attempting to filter the clays, due to vacuum filtration moving rather slowly during separation, clays only were filter for ~ 2.5 hours before actually being placed in the oven.
• The temperature of the oven when the clays were placed in was ~ 90°C.
• When removing clays from freeze and thaw process in cycle #1, many clays appeared to maintain ice crystals within their structure.
• NaMT clays after freezing and thawing cycles appeared more rigid than laponite samples, but all in all PVOH 146K structure was maintained.
• As freezing and thawing continued, PVOH 146K control sample became more crystalline and clear in appearance.
• For all hydrogels that were removed with dye previous placed and attached within their contents, ~ 3mL of water was added to each beaker to analyze as well as quantify if and how much dye would leak out of each hydrogel.
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Friday, June 22, 2007
Who Wouldn't Want Bill Cowher's Tie?
PSAMP HAS MOVED! CLICK HERE TO GET TO THE BRAND NEW SITE! NOW!
Here's a cool fundraiser that you should definitely check out.
Allegheny County Chief Executive Dan Onorato announced the auction of 30 celebrity ties that are available for your online bid. The ties, which have been "touring" Pittsburgh for the past month, will be auctioned online at the County's website.
All proceeds will benefit the Kane Foundation, which raises private funds to help enhance the quality of life of the residents of Allegheny County’s four Kane Regional Centers and their families. The Foundation helps to fund adult education classes, cultural and sporting events, and pet therapy, among other activities.
Want to know more about the Kane Centers and who they serve? Glad you asked.
The Kane Regional Centers offer residential skilled nursing care and rehabilitation for short-term and long-term needs in Glen Hazel, McKeesport, Ross Township and Scott Township. The Glen Hazel and Ross Centers also offer comprehensive living centers with a spectrum of short- and long-term residential and non-residential services.
In short, the assisted living centers help improve the quality of life to the residents of all the centers.
Bids will be taken online at this link. Those with the highest bid will receive the autographed tie, along with a caricature of the former tie-owner. PSaMP would go for the Dan Rooney one. It looks rather snazzy.
Here's a list of the big-timers who graciously donated ties to help this cool cause!
The Honorable Joe Biden, U.S. Senator, Delaware
The Honorable Robert P. Casey, Jr., U.S. Senator, Pennsylvania
Bill Cowher, Former Pittsburgh Steelers Coach
The Honorable Mike Doyle, U.S. Congressman, Pennsylvania
Dr. Freddie Fu, Orthopedic Surgeon & Sports Medicine Expert
The Honorable Ted Kennedy, U.S. Senator, Massachusetts
Jimmy Kimmel, Jimmy Kimmel Live Host
Bobby Knight, Texas Tech Basketball Coach
The Honorable Catherine Baker Knoll, Pennsylvania Lt. Governor
Jimmy Krenn, WDVE Morning Show Host
Jay Leno, Tonight Show Host
Kevin McClatchy, Pittsburgh Pirates Chief Executive Officer
Robert Nutting, Pittsburgh Pirates Chairman
The Honorable Dan Onorato, Allegheny County Chief Executive
The Honorable Mark Pryor, U.S. Senator, Arkansas
The Honorable Luke Ravenstahl, Pittsburgh Mayor
The Honorable Jay Rockefeller, U.S. Senator, West Virginia
Jim Roddey, Former Allegheny County Chief Executive
Dan Rooney, Pittsburgh Steelers Chairman
Tim Russert, Meet the Press Host
Ray Shero, Pittsburgh Penguins General Manager
Max Starks, Pittsburgh Steelers Offensive Lineman
George Steinbrenner, New York Yankees Owner
The Honorable Ted Stevens, U.S. Senator, Alaska
Rod Stewart, Singer
Andrew Stockey, WTAE-TV Anchor
Mike Tomlin, Pittsburgh Steelers Coach
Dave Wannstedt, Pitt Football Coach
The Honorable Jack Wagner, Pennsylvania Auditor General
George Will, National Columnist
Tons of Pittsburgh sports icons on this list. This unique auction will help you get that much closer to the celebrities you know and love.
Not a Pittsburgh sports fan? Why not go for Rod Stewart's tie? Or how about Bobby Knight or Jay Leno?
Online bidding runs through June 30th, so act fast.
Many thanks to Sean for supplying the link to this worthy cause. PSaMP is all for raising money/helping the community, and the Kane Foundation totally fits the bill.
Now you do your part. Bid on some cool ties and help support the Kane Centers!
PSAMP HAS MOVED! CLICK HERE TO GET TO THE BRAND NEW SITE! NOW!
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Through some strange and powerful principle of mental chemistry which she has never divulged, nature wraps up in the impulse of strong desire, that something which recognizes no such word as impossible, and accepts no such reality as failure. Hill, Napoleon
This quote is about desire · Search on Google Books to find all references and sources for this quotation.
A bit about Hill, Napoleon ...
Napoleon Hill (October 26, 1883 November 8, 1970) was an American author who was one the earliest producers of the modern genre of personal-success literature. His most famous work, Think and Grow Rich, is one of the best-selling books of all time.
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A book worth reading is worth buying. Ruskin, John
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A bit about Ruskin, John ...
John Ruskin (February 8, 1819 January 20, 1900) was an English author, poet and artist, although more famous for his work as art critic and social critic. Ruskin's thinking on art and architecture became the thinking of the Victorian and Edwardian eras.
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